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Santa Cruz Biotechnology mmp 2
Comparison of MMP-9 and <t>MMP-2</t> levels in perfusates from kidneys from DBD (n = 15) and cDCDD (n = 9) human donors. Error bars represent Standard Error of the Mean (SEM). * p
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1) Product Images from "Protection of the Transplant Kidney from Preservation Injury by Inhibition of Matrix Metalloproteinases"

Article Title: Protection of the Transplant Kidney from Preservation Injury by Inhibition of Matrix Metalloproteinases

Journal: PLoS ONE

doi: 10.1371/journal.pone.0157508

Comparison of MMP-9 and MMP-2 levels in perfusates from kidneys from DBD (n = 15) and cDCDD (n = 9) human donors. Error bars represent Standard Error of the Mean (SEM). * p
Figure Legend Snippet: Comparison of MMP-9 and MMP-2 levels in perfusates from kidneys from DBD (n = 15) and cDCDD (n = 9) human donors. Error bars represent Standard Error of the Mean (SEM). * p

Techniques Used:

MMP-2 expression in perfusate from rat kidney cold perfused with KPS-1 solution with MMP-2 siRNA added (n = 4). Insert: a representative zymography for MMP-2. Error bars represent Standard Error of the Mean (SEM). * p
Figure Legend Snippet: MMP-2 expression in perfusate from rat kidney cold perfused with KPS-1 solution with MMP-2 siRNA added (n = 4). Insert: a representative zymography for MMP-2. Error bars represent Standard Error of the Mean (SEM). * p

Techniques Used: Expressing, Zymography

Level of undamaged mitochondria measured by the levels of cytochrome c oxidase activity in rat kidney tissue after initial flush and following 22 hours of machine cold perfusion with KPS-1, KPS-1 with doxycycline, and KPS-1 with MMP-2 siRNA (n = 4). Error bars represent Standard Error of the Mean (SEM). * p
Figure Legend Snippet: Level of undamaged mitochondria measured by the levels of cytochrome c oxidase activity in rat kidney tissue after initial flush and following 22 hours of machine cold perfusion with KPS-1, KPS-1 with doxycycline, and KPS-1 with MMP-2 siRNA (n = 4). Error bars represent Standard Error of the Mean (SEM). * p

Techniques Used: Activity Assay

Matrix Metalloproteinase levels and injury markers in perfusate from clinically transplanted kidneys with DGF (n = 5) and without DGF (n = 19). A: MMP-9 level B: MMP-2 level C: NGAL levelD: LDH activity Error bars represent Standard Error of the Mean (SEM). * p
Figure Legend Snippet: Matrix Metalloproteinase levels and injury markers in perfusate from clinically transplanted kidneys with DGF (n = 5) and without DGF (n = 19). A: MMP-9 level B: MMP-2 level C: NGAL levelD: LDH activity Error bars represent Standard Error of the Mean (SEM). * p

Techniques Used: Activity Assay

Effect of doxycycline on LDH, cytochrome c oxidase, and total protein release. A: Confirmation of inhibition of MMP-2 and MMP-9 activity by doxycycline using zymography. B: LDH activity in perfusate of rat kidney with warm ischemic injury at 5, 10, and 22 hours perfused with KPS-1 with and without doxycycline (n = 7 each). C: Cytochrome c oxidase level in perfusate of rat kidney with warm ischemic injury at 5, 10, and 22 hours perfused with KPS-1 with and without doxycycline (n = 7 each). D: Total protein release in perfusate of rat kidney with warm ischemic injury at 5, 10, and 22 hours perfused with KPS-1 with and without doxycycline (n = 6 each). Error bars represent Standard Error of the Mean (SEM). * p
Figure Legend Snippet: Effect of doxycycline on LDH, cytochrome c oxidase, and total protein release. A: Confirmation of inhibition of MMP-2 and MMP-9 activity by doxycycline using zymography. B: LDH activity in perfusate of rat kidney with warm ischemic injury at 5, 10, and 22 hours perfused with KPS-1 with and without doxycycline (n = 7 each). C: Cytochrome c oxidase level in perfusate of rat kidney with warm ischemic injury at 5, 10, and 22 hours perfused with KPS-1 with and without doxycycline (n = 7 each). D: Total protein release in perfusate of rat kidney with warm ischemic injury at 5, 10, and 22 hours perfused with KPS-1 with and without doxycycline (n = 6 each). Error bars represent Standard Error of the Mean (SEM). * p

Techniques Used: Inhibition, Activity Assay, Zymography

Comparison of MMP-9 and MMP-2 levels at 5, 10, and 22 hours in perfusates from rat kidney with warm ischemic injury undergoing machine cold perfusion (n = 7). Insert: representative zymography with MMP-9 and MMP-2 activitiesError bars represent Standard Error of the Mean (SEM). * p
Figure Legend Snippet: Comparison of MMP-9 and MMP-2 levels at 5, 10, and 22 hours in perfusates from rat kidney with warm ischemic injury undergoing machine cold perfusion (n = 7). Insert: representative zymography with MMP-9 and MMP-2 activitiesError bars represent Standard Error of the Mean (SEM). * p

Techniques Used: Zymography

NGAL in perfusate of rat kidney with warm ischemic injury at 22 hours with KPS-1 and KPS-1 supplemented with doxycycline and MMP-2 siRNA compared to control (5 hours) (n = 4). Error bars represent Standard Error of the Mean (SEM). * p
Figure Legend Snippet: NGAL in perfusate of rat kidney with warm ischemic injury at 22 hours with KPS-1 and KPS-1 supplemented with doxycycline and MMP-2 siRNA compared to control (5 hours) (n = 4). Error bars represent Standard Error of the Mean (SEM). * p

Techniques Used:

2) Product Images from "MMP-2 siRNA Inhibits Radiation-Enhanced Invasiveness in Glioma Cells"

Article Title: MMP-2 siRNA Inhibits Radiation-Enhanced Invasiveness in Glioma Cells

Journal: PLoS ONE

doi: 10.1371/journal.pone.0020614

p-MMP-2 combined with radiation enhances apoptosis in vivo . A, Immunohistochemical analysis of brain sections using anti-MMP-2, anti-VEGF and anti-pFAK antibodies. Sections were photographed (60×). Also shown is the negative control where the primary antibody was replaced by non-specific IgG (insets). B, Tissue sections of mice were evaluated with the TUNEL assay according to manufacturer's instructions and photographed under fluorescent microscopy (60×). For the negative control, samples were incubated with label solution (without terminal transferase) instead of TUNEL reaction mixture (insets). C, siRNA against MMP-2 inhibits U251 tumor cell invasion in vivo . H E staining was performed according to standard protocol, and representative pictures of tumor sections from mock, pSV, p-MMP-2-treated mice are shown (20× and 60×). D , Immunohistochemical analysis of brain sections using anti-human nuclei (HuNu) antibody, a histological marker for identification of human cells (a specific human nuclear antigen). Entire brain sections were photographed (4×; middle row); shown on the top row is a non-tumor region (40×; top row); and shown on the bottom row is tumor and non-tumor overlapping region (40×; bottom row). Also shown is the negative control where the primary antibody was replaced by non-specific IgG (inset).
Figure Legend Snippet: p-MMP-2 combined with radiation enhances apoptosis in vivo . A, Immunohistochemical analysis of brain sections using anti-MMP-2, anti-VEGF and anti-pFAK antibodies. Sections were photographed (60×). Also shown is the negative control where the primary antibody was replaced by non-specific IgG (insets). B, Tissue sections of mice were evaluated with the TUNEL assay according to manufacturer's instructions and photographed under fluorescent microscopy (60×). For the negative control, samples were incubated with label solution (without terminal transferase) instead of TUNEL reaction mixture (insets). C, siRNA against MMP-2 inhibits U251 tumor cell invasion in vivo . H E staining was performed according to standard protocol, and representative pictures of tumor sections from mock, pSV, p-MMP-2-treated mice are shown (20× and 60×). D , Immunohistochemical analysis of brain sections using anti-human nuclei (HuNu) antibody, a histological marker for identification of human cells (a specific human nuclear antigen). Entire brain sections were photographed (4×; middle row); shown on the top row is a non-tumor region (40×; top row); and shown on the bottom row is tumor and non-tumor overlapping region (40×; bottom row). Also shown is the negative control where the primary antibody was replaced by non-specific IgG (inset).

Techniques Used: In Vivo, Immunohistochemistry, Negative Control, Mouse Assay, TUNEL Assay, Microscopy, Incubation, Staining, Marker

p-MMP-2 inhibits colony formation and induces apoptosis and activation of pro-apoptotic molecules in glioma cells. A, Clonogenic assay was performed as described in Materials and Methods . The cells were cultured, and colonies larger than 50 cells were counted. Columns : mean of triplicate experiments; bars : SD; * p
Figure Legend Snippet: p-MMP-2 inhibits colony formation and induces apoptosis and activation of pro-apoptotic molecules in glioma cells. A, Clonogenic assay was performed as described in Materials and Methods . The cells were cultured, and colonies larger than 50 cells were counted. Columns : mean of triplicate experiments; bars : SD; * p

Techniques Used: Activation Assay, Clonogenic Assay, Cell Culture

p-MMP-2 transfection inhibits radiation-enhanced glioma cell invasion. U-251 and U-87 cells were transfected with mock, p-SV or p-MMP-2, and irradiated as described earlier. Cells were trypsinized and counted, and 5×10 5 cells from each treatment condition were allowed to invade transwell inserts containing 12-µm-pore polycarbonate membranes pre-coated with Matrigel for 24 h at 37°C. Afterwards, cells were fixed and stained with Hema-3. Cells that had migrated to the lower side of the membrane were photographed under a light microscope at 20× magnification. Percentages of invading cells were quantified by counting five fields from each treatment condition. Columns : mean of triplicate experiments; bars : SD; * p
Figure Legend Snippet: p-MMP-2 transfection inhibits radiation-enhanced glioma cell invasion. U-251 and U-87 cells were transfected with mock, p-SV or p-MMP-2, and irradiated as described earlier. Cells were trypsinized and counted, and 5×10 5 cells from each treatment condition were allowed to invade transwell inserts containing 12-µm-pore polycarbonate membranes pre-coated with Matrigel for 24 h at 37°C. Afterwards, cells were fixed and stained with Hema-3. Cells that had migrated to the lower side of the membrane were photographed under a light microscope at 20× magnification. Percentages of invading cells were quantified by counting five fields from each treatment condition. Columns : mean of triplicate experiments; bars : SD; * p

Techniques Used: Transfection, Irradiation, Staining, Light Microscopy

p-MMP-2 transfection in combination with radiation inhibits glioma cell migration. U-251 and U-87 cells were cultured for formation of spheroids as described in Materials and Methods . Spheroids were then transfected with mock, p-SV or p-MMP-2, and followed by irradiation as described earlier. At the end of the migration assay, spheroids were fixed and stained with Hema-3. Migration of cells from spheroids to monolayers was measured using a microscope calibrated with a stage and ocular micrometer. Columns : mean of triplicate experiments; bars : SD; * p
Figure Legend Snippet: p-MMP-2 transfection in combination with radiation inhibits glioma cell migration. U-251 and U-87 cells were cultured for formation of spheroids as described in Materials and Methods . Spheroids were then transfected with mock, p-SV or p-MMP-2, and followed by irradiation as described earlier. At the end of the migration assay, spheroids were fixed and stained with Hema-3. Migration of cells from spheroids to monolayers was measured using a microscope calibrated with a stage and ocular micrometer. Columns : mean of triplicate experiments; bars : SD; * p

Techniques Used: Transfection, Migration, Cell Culture, Irradiation, Staining, Microscopy

p-MMP-2 combined with radiation inhibits tumor growth in vivo . U-251 (1×10 6 ) cells were injected intracerebrally into athymic mice. After ten days, animals were separated into five groups and were treated on alternate days with intracerebral injections of p-SV or p-MMP-2 for a total of 4 doses (60 µg per dose) and 2 doses of radiation (4 Gy per dose) as described in Materials and Methods . Six weeks after the experiment was initiated, mice were euthanized with intracardiac perfusion of PBS, followed by formaldehyde. The brains were then removed. A, Six weeks after the experiment was initiated, an intraperitonal injection of 2.5 mg D-luciferin sodium salt diluted in 50 µL of PBS was given, and animals were photographed under the IVIS camera for fluorescent light emission. The brains were removed and fixed in 10% phosphate-buffered formaldehyde, and the fixed tissue samples were then processed into paraffin blocks. Brain sections (5 µM thick) were stained with hematoxylin and eosin (H E), and photographed under a light microscope (4× and 40×). B, Every fifth or sixth brain section (5 µM thick) was stained with H E solution, and the tumor masses (H E-stained) were manually traced on the microscope attached computer screen. Areas were calculated using Image Pro Discovery Program software (Media Cybernetics, Inc., Silver Spring, MD). The total tumor volume was calculated as the summed area on all slices, multiplied by the slice separation. Columns : mean of area of tumor portion of all mice in the group (n = 8); bars : SD; * p
Figure Legend Snippet: p-MMP-2 combined with radiation inhibits tumor growth in vivo . U-251 (1×10 6 ) cells were injected intracerebrally into athymic mice. After ten days, animals were separated into five groups and were treated on alternate days with intracerebral injections of p-SV or p-MMP-2 for a total of 4 doses (60 µg per dose) and 2 doses of radiation (4 Gy per dose) as described in Materials and Methods . Six weeks after the experiment was initiated, mice were euthanized with intracardiac perfusion of PBS, followed by formaldehyde. The brains were then removed. A, Six weeks after the experiment was initiated, an intraperitonal injection of 2.5 mg D-luciferin sodium salt diluted in 50 µL of PBS was given, and animals were photographed under the IVIS camera for fluorescent light emission. The brains were removed and fixed in 10% phosphate-buffered formaldehyde, and the fixed tissue samples were then processed into paraffin blocks. Brain sections (5 µM thick) were stained with hematoxylin and eosin (H E), and photographed under a light microscope (4× and 40×). B, Every fifth or sixth brain section (5 µM thick) was stained with H E solution, and the tumor masses (H E-stained) were manually traced on the microscope attached computer screen. Areas were calculated using Image Pro Discovery Program software (Media Cybernetics, Inc., Silver Spring, MD). The total tumor volume was calculated as the summed area on all slices, multiplied by the slice separation. Columns : mean of area of tumor portion of all mice in the group (n = 8); bars : SD; * p

Techniques Used: In Vivo, Injection, Mouse Assay, Staining, Light Microscopy, Microscopy, Software

Radiation enhances MMP-2 and p-MMP-2 inhibits MMP-2 activity and expression in glioma cell lines. A, U-251 and U-87 cells were irradiated with 0–12 Gy X-ray, incubated for 24 h, and conditioned medium collected. MMP-2 activity was determined by gelatin zymography. The band intensities of MMP-2 activity were quantified by densitometry. Columns : mean of triplicate experiments; bars : SD; * p
Figure Legend Snippet: Radiation enhances MMP-2 and p-MMP-2 inhibits MMP-2 activity and expression in glioma cell lines. A, U-251 and U-87 cells were irradiated with 0–12 Gy X-ray, incubated for 24 h, and conditioned medium collected. MMP-2 activity was determined by gelatin zymography. The band intensities of MMP-2 activity were quantified by densitometry. Columns : mean of triplicate experiments; bars : SD; * p

Techniques Used: Activity Assay, Expressing, Irradiation, Incubation, Zymography

p-MMP-2 transfection inhibits radiation-enhanced MMP-2 activity and expression levels as well as cell viability. A, U-251 and U-87 cells were transfected with mock (PBS), p-SV or p-MMP-2 (2 µg), and after 72 h of incubation, cells were irradiated with 0, 2, 4, 6 or 8 Gy and incubated for a further 24 h. Conditioned media was used to determine MMP-2 activity by gelatin zymography, and total cell lysates were used to determine MMP-2 levels by Western blotting. B, Total RNA was used to determine MMP-2 mRNA transcription levels by RT-PCR with gene-specific primers. GAPDH served as a loading control. C, U-251 and U-87 cells were transfected with mock, p-SV or p-MMP-2 and irradiated as described above. 24 h after radiation, the cells were fixed and processed to visualize MMP-2 expression. The cells were mounted using mounting media with DAPI to visualize the nucleus. D, U-251 and U-87 cells were transfected with mock, p-SV or p-MMP-2, and irradiated for 72 h after transfection. After a another 24 h of incubation, cell viability was analyzed by MTT assay (absorbance read at 550 nm). Columns : mean of triplicate experiments; bars : SD; * p
Figure Legend Snippet: p-MMP-2 transfection inhibits radiation-enhanced MMP-2 activity and expression levels as well as cell viability. A, U-251 and U-87 cells were transfected with mock (PBS), p-SV or p-MMP-2 (2 µg), and after 72 h of incubation, cells were irradiated with 0, 2, 4, 6 or 8 Gy and incubated for a further 24 h. Conditioned media was used to determine MMP-2 activity by gelatin zymography, and total cell lysates were used to determine MMP-2 levels by Western blotting. B, Total RNA was used to determine MMP-2 mRNA transcription levels by RT-PCR with gene-specific primers. GAPDH served as a loading control. C, U-251 and U-87 cells were transfected with mock, p-SV or p-MMP-2 and irradiated as described above. 24 h after radiation, the cells were fixed and processed to visualize MMP-2 expression. The cells were mounted using mounting media with DAPI to visualize the nucleus. D, U-251 and U-87 cells were transfected with mock, p-SV or p-MMP-2, and irradiated for 72 h after transfection. After a another 24 h of incubation, cell viability was analyzed by MTT assay (absorbance read at 550 nm). Columns : mean of triplicate experiments; bars : SD; * p

Techniques Used: Transfection, Activity Assay, Expressing, Incubation, Irradiation, Zymography, Western Blot, Reverse Transcription Polymerase Chain Reaction, MTT Assay

p-MMP-2 inhibits radiation-enhanced tumor culture medium-induced microtubule network formation in endothelial cells and downregulates expression of angiogenesis-associated molecules. A, Human microvascular endothelial cells (5×10 4 ) were seeded in 96-well plates and cultured with conditioned medium collected from U-251 and U-87 glioma cells transfected with mock, p-SV, and p-MMP-2, and irradiated as described earlier. 24 h after radiation treatment, the cells were washed, fixed and stained with Hema-3 and photographed. Percentages of branches were quantified by counting five fields in each condition. Columns : mean of triplicate experiments; bars : SD; * p
Figure Legend Snippet: p-MMP-2 inhibits radiation-enhanced tumor culture medium-induced microtubule network formation in endothelial cells and downregulates expression of angiogenesis-associated molecules. A, Human microvascular endothelial cells (5×10 4 ) were seeded in 96-well plates and cultured with conditioned medium collected from U-251 and U-87 glioma cells transfected with mock, p-SV, and p-MMP-2, and irradiated as described earlier. 24 h after radiation treatment, the cells were washed, fixed and stained with Hema-3 and photographed. Percentages of branches were quantified by counting five fields in each condition. Columns : mean of triplicate experiments; bars : SD; * p

Techniques Used: Expressing, Cell Culture, Transfection, Irradiation, Staining

3) Product Images from "MMP-2 Alters VEGF Expression via ?V?3 Integrin-Mediated PI3K/AKT Signaling in A549 Lung Cancer Cells"

Article Title: MMP-2 Alters VEGF Expression via ?V?3 Integrin-Mediated PI3K/AKT Signaling in A549 Lung Cancer Cells

Journal: International journal of cancer. Journal international du cancer

doi: 10.1002/ijc.25134

Recombinant human-MMP-2 inhibits Ad-MMP-2-Si-inhibited PI3K, phosphorylation of AKT and VEGF expression in lung adenocarcinoma cells (A B) After 24 h of Ad-MMP-2-Si infection, A549/H1299 cells were incubated with rhMMP-2 (25 ng/mL) for 12 h and the medium was aspirated, cells were washed with PBS three times, a minimum amount of serum-free medium was added, and cells were incubated for another 12 h. We then performed western blot analysis for VEGF in conditioned medium (CM) and expression of PI3K, phosphorylated-AKT (Ser-473), VEGF and VEGFR-2 in cell lysates (CL). Protein band intensities were quantified by densitometric analysis using ImageJ software (National Institutes of Health). The levels of PI3K, phospho-AKT (Ser-473) and VEGF proteins were normalized to respective protein level in mock infected cells. Columns : mean of triplicate experiments; ** p
Figure Legend Snippet: Recombinant human-MMP-2 inhibits Ad-MMP-2-Si-inhibited PI3K, phosphorylation of AKT and VEGF expression in lung adenocarcinoma cells (A B) After 24 h of Ad-MMP-2-Si infection, A549/H1299 cells were incubated with rhMMP-2 (25 ng/mL) for 12 h and the medium was aspirated, cells were washed with PBS three times, a minimum amount of serum-free medium was added, and cells were incubated for another 12 h. We then performed western blot analysis for VEGF in conditioned medium (CM) and expression of PI3K, phosphorylated-AKT (Ser-473), VEGF and VEGFR-2 in cell lysates (CL). Protein band intensities were quantified by densitometric analysis using ImageJ software (National Institutes of Health). The levels of PI3K, phospho-AKT (Ser-473) and VEGF proteins were normalized to respective protein level in mock infected cells. Columns : mean of triplicate experiments; ** p

Techniques Used: Recombinant, Expressing, Infection, Incubation, Western Blot, Software

MMP-2 inhibition using Ad-MMP-2-Si decreased angiogenesis, VEGF expression, and co-localization of MMP-2 and integrin-αVβ3 in A549 lung tumors in vivo H E staining was performed as per standard protocol and representative pictures (400 X magnifications) of tumor sections from mock, Ad-SV and Ad-MMP-2-Si-treated mice were shown. Immunohistochemistry was performed for expression and co-localization (yellow) of MMP-2 (green) and integrin-αVβ3 (Red) using specific antibodies. Also showed immunohistochemical analysis for anti-Human Von-Willebrand Factor (Factor-VIII) and VEGF expression using specific antibodies. Data shown are representative fields (400 X magnifications). Also shown is the negative control where the primary antibody was replaced by non-immune serum (inserts).
Figure Legend Snippet: MMP-2 inhibition using Ad-MMP-2-Si decreased angiogenesis, VEGF expression, and co-localization of MMP-2 and integrin-αVβ3 in A549 lung tumors in vivo H E staining was performed as per standard protocol and representative pictures (400 X magnifications) of tumor sections from mock, Ad-SV and Ad-MMP-2-Si-treated mice were shown. Immunohistochemistry was performed for expression and co-localization (yellow) of MMP-2 (green) and integrin-αVβ3 (Red) using specific antibodies. Also showed immunohistochemical analysis for anti-Human Von-Willebrand Factor (Factor-VIII) and VEGF expression using specific antibodies. Data shown are representative fields (400 X magnifications). Also shown is the negative control where the primary antibody was replaced by non-immune serum (inserts).

Techniques Used: Inhibition, Expressing, In Vivo, Staining, Mouse Assay, Immunohistochemistry, Negative Control

Ad-MMP-2-Si inhibits tumor cell conditioned medium-induced angiogenesis in HMEC-1 cells and inhibits VEGF and VEGFR-2 expression in A549 cells (A) In vitro angiogenesis: A549 cells were infected with mock, Ad-SV or Ad-MMP-2-Si for 36 h, medium was removed, minimum amounts of serum-free medium were added to cover the cells, and cells were incubated for 12 h. Tumor conditioned medium (TCM) was collected and added into 96-well plates, which were coated with Matrigel and seeded with human dermal microvascular endothelial cells-1 (HMEC-1; 2×10 4 cells/well). After overnight incubation at 37°C, cells were observed under the bright field microscope for formation of capillary-like structures. The degree of angiogenic induction by mock-TCM, Ad-SV-TCM and Ad-MMP-2-Si-TCM was quantified for the numerical value of the product of the relative capillary length per microscopic field. Columns : mean of triplicate experiments; bars : SE; * p
Figure Legend Snippet: Ad-MMP-2-Si inhibits tumor cell conditioned medium-induced angiogenesis in HMEC-1 cells and inhibits VEGF and VEGFR-2 expression in A549 cells (A) In vitro angiogenesis: A549 cells were infected with mock, Ad-SV or Ad-MMP-2-Si for 36 h, medium was removed, minimum amounts of serum-free medium were added to cover the cells, and cells were incubated for 12 h. Tumor conditioned medium (TCM) was collected and added into 96-well plates, which were coated with Matrigel and seeded with human dermal microvascular endothelial cells-1 (HMEC-1; 2×10 4 cells/well). After overnight incubation at 37°C, cells were observed under the bright field microscope for formation of capillary-like structures. The degree of angiogenic induction by mock-TCM, Ad-SV-TCM and Ad-MMP-2-Si-TCM was quantified for the numerical value of the product of the relative capillary length per microscopic field. Columns : mean of triplicate experiments; bars : SE; * p

Techniques Used: Expressing, In Vitro, Infection, Incubation, Microscopy

Ad-MMP-2-Si inhibits HIF-1α expression in A549 cells A549 cells were infected with mock, Ad-SV or Ad-MMP-2-Si for 36 h, medium was removed, minimum amounts of serum-free medium were added to cover the cells, and cells were incubated at 1.0% O 2 for 12 h. Finally, we collected the conditioned medium and the cells. (A) Cell lysates were used for western blot analysis for HIF-1α expression using a HIF-1α-specific antibody. The experiments were carried out three times and a representative western blot is shown. The blots were stripped and re-probed with GAPDH antibody to detect total amounts of the respective proteins. Protein band intensities were quantified by densitometric analysis using ImageJ software (National Institutes of Health). The levels of HIF-1α protein were normalized to protein level in mock infected A549 cells. Columns : mean of triplicate experiments; bars : SE; * p
Figure Legend Snippet: Ad-MMP-2-Si inhibits HIF-1α expression in A549 cells A549 cells were infected with mock, Ad-SV or Ad-MMP-2-Si for 36 h, medium was removed, minimum amounts of serum-free medium were added to cover the cells, and cells were incubated at 1.0% O 2 for 12 h. Finally, we collected the conditioned medium and the cells. (A) Cell lysates were used for western blot analysis for HIF-1α expression using a HIF-1α-specific antibody. The experiments were carried out three times and a representative western blot is shown. The blots were stripped and re-probed with GAPDH antibody to detect total amounts of the respective proteins. Protein band intensities were quantified by densitometric analysis using ImageJ software (National Institutes of Health). The levels of HIF-1α protein were normalized to protein level in mock infected A549 cells. Columns : mean of triplicate experiments; bars : SE; * p

Techniques Used: Expressing, Infection, Incubation, Western Blot, Software

MMP-2 interaction with integrin-αVβ3 induces PI3K/AKT-mediated VEGF expression resulting in angiogenesis MMP-2 siRNA inhibits PI3K/AKT-mediated VEGF expression resulting in decreased angiogenesis.
Figure Legend Snippet: MMP-2 interaction with integrin-αVβ3 induces PI3K/AKT-mediated VEGF expression resulting in angiogenesis MMP-2 siRNA inhibits PI3K/AKT-mediated VEGF expression resulting in decreased angiogenesis.

Techniques Used: Expressing

Inhibition of MMP-2 inhibits integrin-αVβ3-mediated VEGF expression in A549 cells (A) A549 cells were plated in 8-well chamber slides and infected with mock or 100 MOI of either Ad-SV or Ad-MMP-2-Si for 48 h. Immunocytochemistry was performed for co-localization (yellow) of MMP-2 (green) and integrin-αVβ3 (Red) using specific antibodies. (B) A549 cells were infected with mock or 100 MOI of either Ad-SV or Ad-MMP-2-Si for 48 h, and proteins were immunoprecipitated with an anti-integrin-αVβ3 antibody. Western blot analysis was performed for MMP-2 using immunoprecipitated proteins. Protein band intensities were quantified by densitometric analysis using ImageJ software (National Institutes of Health). The levels of MMP-2 protein were normalized to protein level in mock infected A549 cells. Columns : mean of triplicate experiments; * p
Figure Legend Snippet: Inhibition of MMP-2 inhibits integrin-αVβ3-mediated VEGF expression in A549 cells (A) A549 cells were plated in 8-well chamber slides and infected with mock or 100 MOI of either Ad-SV or Ad-MMP-2-Si for 48 h. Immunocytochemistry was performed for co-localization (yellow) of MMP-2 (green) and integrin-αVβ3 (Red) using specific antibodies. (B) A549 cells were infected with mock or 100 MOI of either Ad-SV or Ad-MMP-2-Si for 48 h, and proteins were immunoprecipitated with an anti-integrin-αVβ3 antibody. Western blot analysis was performed for MMP-2 using immunoprecipitated proteins. Protein band intensities were quantified by densitometric analysis using ImageJ software (National Institutes of Health). The levels of MMP-2 protein were normalized to protein level in mock infected A549 cells. Columns : mean of triplicate experiments; * p

Techniques Used: Inhibition, Expressing, Infection, Immunocytochemistry, Immunoprecipitation, Western Blot, Software

Ad-MMP-2-Si infection inhibits MMP-2 expression in A549 cells A549 cells were infected as described in Materials and Methods. Briefly, A549 cells were infected with 100 MOI of either Ad-SV or Ad-MMP-2-Si, the medium was aspirated after 36 h of incubation, 3 mL of serum-free medium was added, and cells were incubated overnight. (A) Gelatin zymographic analysis for secreted MMP-2 activity in to the tumor conditioned medium. The gelatinolytic band intensities of MMP-2 were quantified by densitometric analysis using ImageJ software (National Institutes of Health) and normalized with the intensity of the gelatinolytic band in mock-conditioned medium. Columns : mean of triplicate experiments; bars : SE (Standard Error); * p
Figure Legend Snippet: Ad-MMP-2-Si infection inhibits MMP-2 expression in A549 cells A549 cells were infected as described in Materials and Methods. Briefly, A549 cells were infected with 100 MOI of either Ad-SV or Ad-MMP-2-Si, the medium was aspirated after 36 h of incubation, 3 mL of serum-free medium was added, and cells were incubated overnight. (A) Gelatin zymographic analysis for secreted MMP-2 activity in to the tumor conditioned medium. The gelatinolytic band intensities of MMP-2 were quantified by densitometric analysis using ImageJ software (National Institutes of Health) and normalized with the intensity of the gelatinolytic band in mock-conditioned medium. Columns : mean of triplicate experiments; bars : SE (Standard Error); * p

Techniques Used: Infection, Expressing, Incubation, Activity Assay, Software

4) Product Images from "Rho GDIβ promotes Sp1/ MMP‐2 expression and bladder cancer invasion through perturbing miR‐200c‐targeted JNK2 protein translation"

Article Title: Rho GDIβ promotes Sp1/ MMP‐2 expression and bladder cancer invasion through perturbing miR‐200c‐targeted JNK2 protein translation

Journal: Molecular Oncology

doi: 10.1002/1878-0261.12132

JNK 2 specifically mediated Sp1 expression, in turn enhancing mmp‐2 expression and BC cell invasion. (A) The cell extracts obtained from the transfectants, as indicated, were subjected to western blot for determination of protein expression. (B,C) IHC ‐P was carried out to evaluate JNK 2 protein expression in mouse BC tissues as compared with normal bladder tissues. The optical density was analyzed and calculated, as described in Materials and Methods ( n = 12). *Significant difference between the two groups of mice ( P
Figure Legend Snippet: JNK 2 specifically mediated Sp1 expression, in turn enhancing mmp‐2 expression and BC cell invasion. (A) The cell extracts obtained from the transfectants, as indicated, were subjected to western blot for determination of protein expression. (B,C) IHC ‐P was carried out to evaluate JNK 2 protein expression in mouse BC tissues as compared with normal bladder tissues. The optical density was analyzed and calculated, as described in Materials and Methods ( n = 12). *Significant difference between the two groups of mice ( P

Techniques Used: Expressing, Western Blot, Immunohistochemistry, Mouse Assay

Rho GDI β upregulated Sp1 expression and Sp1‐dependent transcription activity in human BC cells. (A) Potential transcription factor binding sites in human mmp‐2 promoter region. (B,C) The cell extracts obtained from (B) T24T(Vector) vs . T24T(Rho GDI β‐ GFP ) cells or (C) T24(Nonsense) vs . T24(shRho GDI β) cells were analyzed for the activation and expression of the transcription factors as indicated. (D,E) The Sp1‐dependent transcriptional activity was evaluated using Sp1‐dependent luciferase reporter. co‐transfected together with pRL ‐ TK into (D) T24T(Vector) and T24T(Rho GDI β‐ GFP ) cells or (E) T24(Nonsense) and T24(shRho GDI β) cells (E). Twenty‐four hours post‐transfection, the transfectants were extracted to evaluate the luciferase activity. TK was used as an internal control. The results are presented as Sp1‐dependent activity relative to the control vector transfectant, and each bar indicates mean ± SD from three independent experiments. *Significant difference ( P
Figure Legend Snippet: Rho GDI β upregulated Sp1 expression and Sp1‐dependent transcription activity in human BC cells. (A) Potential transcription factor binding sites in human mmp‐2 promoter region. (B,C) The cell extracts obtained from (B) T24T(Vector) vs . T24T(Rho GDI β‐ GFP ) cells or (C) T24(Nonsense) vs . T24(shRho GDI β) cells were analyzed for the activation and expression of the transcription factors as indicated. (D,E) The Sp1‐dependent transcriptional activity was evaluated using Sp1‐dependent luciferase reporter. co‐transfected together with pRL ‐ TK into (D) T24T(Vector) and T24T(Rho GDI β‐ GFP ) cells or (E) T24(Nonsense) and T24(shRho GDI β) cells (E). Twenty‐four hours post‐transfection, the transfectants were extracted to evaluate the luciferase activity. TK was used as an internal control. The results are presented as Sp1‐dependent activity relative to the control vector transfectant, and each bar indicates mean ± SD from three independent experiments. *Significant difference ( P

Techniques Used: Expressing, Activity Assay, Binding Assay, Activation Assay, Luciferase, Transfection, Plasmid Preparation

MMP ‐2‐mediated Rho GDI β promoting  BC  invasion. (A–C) Rho GDI β and  MMP ‐2 expressions were analyzed by western blot ( WB ) in (A) T24T(Rho GDI β‐ GFP )  vs . T24T(Vector) cells, (B) T24(shRho GDI β)  vs . T24(Nonsense) and (C) T24T(Vector)  vs . T24T(Rho GDI β‐ GFP /Nonsense) and T24T(Rho GDI β‐ GFP /sh MPP ‐2). (D, E) Invasive abilities of T24T(RhoGDIβ‐GFP/sh MMP ‐2) cells and T24T(Rho GDI β‐ GFP /Nonsense) were determined using  BD  BioCoat™Matrigel™ Invasion Chamber (D), and the relative invasion activity in these two transfectants was plotted (E). *Significant inhibition of invasion in comparison with T24T(Rho GDI β‐ GFP /Nonsense) cells ( P
Figure Legend Snippet: MMP ‐2‐mediated Rho GDI β promoting BC invasion. (A–C) Rho GDI β and MMP ‐2 expressions were analyzed by western blot ( WB ) in (A) T24T(Rho GDI β‐ GFP ) vs . T24T(Vector) cells, (B) T24(shRho GDI β) vs . T24(Nonsense) and (C) T24T(Vector) vs . T24T(Rho GDI β‐ GFP /Nonsense) and T24T(Rho GDI β‐ GFP /sh MPP ‐2). (D, E) Invasive abilities of T24T(RhoGDIβ‐GFP/sh MMP ‐2) cells and T24T(Rho GDI β‐ GFP /Nonsense) were determined using BD BioCoat™Matrigel™ Invasion Chamber (D), and the relative invasion activity in these two transfectants was plotted (E). *Significant inhibition of invasion in comparison with T24T(Rho GDI β‐ GFP /Nonsense) cells ( P

Techniques Used: Western Blot, Activity Assay, Inhibition

miR200c targeted  jnk2  3′‐ UTR , consequently resulting in blockage of  JNK 2 protein translation and inhibition of invasion of  BC  cells. (A) Real‐time  PCR  was used to identify the miR‐200c expression in T24T(Rho GDI β/miR‐200c) cells in comparison with T24T (Rho GDI β/Vector) cells. (B) The cell extracts from T24T(Rho GDI β/miR‐200c) and T24T(Rho GDI β/Vector) cells were used to evaluate the effect of miR‐200c on expression of  JNK 1,  JNK 2, Sp1, and  MMP ‐2.  GAPDH  was used as a protein loading control. (C) The  jnk 2 3′‐ UTR  activity was evaluated by transfection of  jnk2  3′‐ UTR ‐driven luciferase reporter together with  pRL ‐ TK  into the transfectants as indicated. The bars indicate mean ±  SD  from three independent experiments. *Significant difference ( P 
Figure Legend Snippet: miR200c targeted jnk2 3′‐ UTR , consequently resulting in blockage of JNK 2 protein translation and inhibition of invasion of BC cells. (A) Real‐time PCR was used to identify the miR‐200c expression in T24T(Rho GDI β/miR‐200c) cells in comparison with T24T (Rho GDI β/Vector) cells. (B) The cell extracts from T24T(Rho GDI β/miR‐200c) and T24T(Rho GDI β/Vector) cells were used to evaluate the effect of miR‐200c on expression of JNK 1, JNK 2, Sp1, and MMP ‐2. GAPDH was used as a protein loading control. (C) The jnk 2 3′‐ UTR activity was evaluated by transfection of jnk2 3′‐ UTR ‐driven luciferase reporter together with pRL ‐ TK into the transfectants as indicated. The bars indicate mean ±  SD from three independent experiments. *Significant difference ( P 

Techniques Used: Inhibition, Real-time Polymerase Chain Reaction, Expressing, Plasmid Preparation, Activity Assay, Transfection, Luciferase

The gene mmp‐2 was upregulated at the transcriptional level by Rho GDI β. (A–F) Total RNA was extracted from the cells, as indicated. After reverse transcription, the mRNA levels of mmp‐2 in cells were evaluated using RT ‐ PCR assay (A, B) and real‐time PCR assay (C, D). mmp‐2 mRNA stabilities were evaluated by regular RT ‐ PCR (E) or real‐time PCR (F) in the presence of Act D in both T24T(Vector) and T24T(RhoGDIβ‐GFP) cells. (G, H) Wild‐type mmp‐2 promoter‐driven luciferase reporter was co‐transfected together with pRL ‐ TK into T24T(Vector) and T24T(RhoGDIβ‐GFP) cells (G), or T24(Nonsense) and T24(shRho GDI β) cells (H), respectively. Twenty‐four hours post‐transfection, the luciferase activity was evaluated. TK was used as internal control. The results are presented as mmp‐2 promoter activity relative to the control vector transfectant, and each bar indicates mean ± SD from three independent experiments. *Significant increase ( P
Figure Legend Snippet: The gene mmp‐2 was upregulated at the transcriptional level by Rho GDI β. (A–F) Total RNA was extracted from the cells, as indicated. After reverse transcription, the mRNA levels of mmp‐2 in cells were evaluated using RT ‐ PCR assay (A, B) and real‐time PCR assay (C, D). mmp‐2 mRNA stabilities were evaluated by regular RT ‐ PCR (E) or real‐time PCR (F) in the presence of Act D in both T24T(Vector) and T24T(RhoGDIβ‐GFP) cells. (G, H) Wild‐type mmp‐2 promoter‐driven luciferase reporter was co‐transfected together with pRL ‐ TK into T24T(Vector) and T24T(RhoGDIβ‐GFP) cells (G), or T24(Nonsense) and T24(shRho GDI β) cells (H), respectively. Twenty‐four hours post‐transfection, the luciferase activity was evaluated. TK was used as internal control. The results are presented as mmp‐2 promoter activity relative to the control vector transfectant, and each bar indicates mean ± SD from three independent experiments. *Significant increase ( P

Techniques Used: Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Activated Clotting Time Assay, Luciferase, Transfection, Activity Assay, Plasmid Preparation

Sp1 contributed to mmp‐2 transcription and further invasion of BC cells. (A) Illustration of the mutated Sp1 binding site in the mmp‐2 promoter region. (B,C) Wild‐type or mutant mmp ‐2 promoter‐driven luciferase reporters was co‐transfected together with pRL‐TK into (B) T24T(Vector) and T24T(RhoGDIβ‐GFP) cells or (C) T24(Nonsense) and T24(shRhoGDIβ) cells. Twenty‐four hours post‐transfection, the transfectants were extracted to evaluate the luciferase activity. The results were presented as mmp‐2 promoter activity relative to the scramble vector transfectant transfected with wild‐type mmp‐2 promoter‐driven luciferase reporter, and each bar indicates mean ± SD from three independent experiments. *Significant difference in comparison to scramble vector transfetant. ♣Significant difference in comparison with wild‐type reporter transfectant ( P
Figure Legend Snippet: Sp1 contributed to mmp‐2 transcription and further invasion of BC cells. (A) Illustration of the mutated Sp1 binding site in the mmp‐2 promoter region. (B,C) Wild‐type or mutant mmp ‐2 promoter‐driven luciferase reporters was co‐transfected together with pRL‐TK into (B) T24T(Vector) and T24T(RhoGDIβ‐GFP) cells or (C) T24(Nonsense) and T24(shRhoGDIβ) cells. Twenty‐four hours post‐transfection, the transfectants were extracted to evaluate the luciferase activity. The results were presented as mmp‐2 promoter activity relative to the scramble vector transfectant transfected with wild‐type mmp‐2 promoter‐driven luciferase reporter, and each bar indicates mean ± SD from three independent experiments. *Significant difference in comparison to scramble vector transfetant. ♣Significant difference in comparison with wild‐type reporter transfectant ( P

Techniques Used: Binding Assay, Mutagenesis, Luciferase, Transfection, Activity Assay, Plasmid Preparation

5) Product Images from "Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells"

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells

Journal: Acta Biochimica et Biophysica Sinica

doi: 10.1093/abbs/gmw030

ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression
Figure Legend Snippet: ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression

Techniques Used: Over Expression, Western Blot, Stable Transfection, Expressing

Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC
Figure Legend Snippet: Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC

Techniques Used: Expressing

Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly
Figure Legend Snippet: Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly

Techniques Used: Immunohistochemistry, Staining

Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells
Figure Legend Snippet: Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells

Techniques Used: Over Expression, Expressing, Activation Assay, Western Blot

Expressions of Aurora-A and MMP-2 proteins in ESCC tissue
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins in ESCC tissue

Techniques Used:

Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and

Techniques Used: Expressing, Western Blot

6) Product Images from "Expression analysis and clinical significance of eIF4E, VEGF-C, E-cadherin and MMP-2 in colorectal adenocarcinoma"

Article Title: Expression analysis and clinical significance of eIF4E, VEGF-C, E-cadherin and MMP-2 in colorectal adenocarcinoma

Journal: Oncotarget

doi: 10.18632/oncotarget.13453

RL bioluminescence from HCT-15/Rluc cells present in Lung tissue in living mice A. The HCT-15/Rluc/eIF4E cells (1.0 × 10 6 ) and corresponding HCT-15-Rluc were injected via tail-vein 3 days later. The bioluminescence seen represents the thorax region of the mouse where HCT-15/Rluc and HCT-15/Rluc/eIF4E cells are trapped in the lungs. B. The HCT-15/Rluc/VEGF-C cells (1.0 × 10 6 ) and corresponding HCT-15/Rluc were injected via tail-vein 3 days later. The bioluminescence seen represents the thorax region of the mouse where HCT-15/Rluc and HCT-15/Rluc/VEGF-C cells are trapped in the lungs. C. The HCT-15/Rluc/MMP-2 cells (1.0 × 10 6 ) and corresponding HCT-15/Rluc were injected via tail-vein 3 days later. The bioluminescence seen represents the thorax region of the mouse where HCT-15/Rluc and HCT-15/Rluc/MMP-2 cells are trapped in the lungs. D. The HCT/15-Rluc/E-cadherin cells (1.0 × 10 6 ) and corresponding HCT-15-Rluc were injected via tail-vein 3 days later. The bioluminescence seen represents the thorax region of the mouse where HCT-15/Rluc and HCT-15/Rluc/E-cadherin cells are trapped in the lungs.
Figure Legend Snippet: RL bioluminescence from HCT-15/Rluc cells present in Lung tissue in living mice A. The HCT-15/Rluc/eIF4E cells (1.0 × 10 6 ) and corresponding HCT-15-Rluc were injected via tail-vein 3 days later. The bioluminescence seen represents the thorax region of the mouse where HCT-15/Rluc and HCT-15/Rluc/eIF4E cells are trapped in the lungs. B. The HCT-15/Rluc/VEGF-C cells (1.0 × 10 6 ) and corresponding HCT-15/Rluc were injected via tail-vein 3 days later. The bioluminescence seen represents the thorax region of the mouse where HCT-15/Rluc and HCT-15/Rluc/VEGF-C cells are trapped in the lungs. C. The HCT-15/Rluc/MMP-2 cells (1.0 × 10 6 ) and corresponding HCT-15/Rluc were injected via tail-vein 3 days later. The bioluminescence seen represents the thorax region of the mouse where HCT-15/Rluc and HCT-15/Rluc/MMP-2 cells are trapped in the lungs. D. The HCT/15-Rluc/E-cadherin cells (1.0 × 10 6 ) and corresponding HCT-15-Rluc were injected via tail-vein 3 days later. The bioluminescence seen represents the thorax region of the mouse where HCT-15/Rluc and HCT-15/Rluc/E-cadherin cells are trapped in the lungs.

Techniques Used: Mouse Assay, Injection

The expression of eIF4E, VEGF-C, MMP-2 and E-cadherin in primary site of patient-derived colon cancer xenografts A. The negative expression of eIF4E in colon cancerous tissues. B. The positive expression of eIF4E in colon cancerous tissues. C. The negative expression of VEGF-C in colon cancerous tissues. D. The positive expression of VEGF-C in colon cancerous tissues. E. The negative expression of E-cadherin in colon cancerous tissues. F. The positive expression of E-cadherin in colon cancerous tissues. G. The negative expression of MMP-2 in colon cancerous tissues. H. The positive expression of MMP-2 in colon cancerous tissues. Bar = 25 μM.
Figure Legend Snippet: The expression of eIF4E, VEGF-C, MMP-2 and E-cadherin in primary site of patient-derived colon cancer xenografts A. The negative expression of eIF4E in colon cancerous tissues. B. The positive expression of eIF4E in colon cancerous tissues. C. The negative expression of VEGF-C in colon cancerous tissues. D. The positive expression of VEGF-C in colon cancerous tissues. E. The negative expression of E-cadherin in colon cancerous tissues. F. The positive expression of E-cadherin in colon cancerous tissues. G. The negative expression of MMP-2 in colon cancerous tissues. H. The positive expression of MMP-2 in colon cancerous tissues. Bar = 25 μM.

Techniques Used: Expressing, Derivative Assay

The growth rates of colon cancer cell lines HCT-15/Rluc with overexpression of E-cadherin, MMP-2, VEGF-c and eIF4E and parental HCT-15/Rluc A. Western blotting showed that the stable cell lines including HCT-15/Rluc/E-cadherin, HCT-15/Rluc/VEGF-C, HCT-15/Rluc/MMP-2, and HCT-15/Rluc/eIF4E were constructed successfully. B. The stable colon cancer cell lines with overexpression of E-cadherin, MMP-2, VEGF-c and eIF4E as well as parental HCT-15/Rluc cell line showed different growth rates.
Figure Legend Snippet: The growth rates of colon cancer cell lines HCT-15/Rluc with overexpression of E-cadherin, MMP-2, VEGF-c and eIF4E and parental HCT-15/Rluc A. Western blotting showed that the stable cell lines including HCT-15/Rluc/E-cadherin, HCT-15/Rluc/VEGF-C, HCT-15/Rluc/MMP-2, and HCT-15/Rluc/eIF4E were constructed successfully. B. The stable colon cancer cell lines with overexpression of E-cadherin, MMP-2, VEGF-c and eIF4E as well as parental HCT-15/Rluc cell line showed different growth rates.

Techniques Used: Over Expression, Western Blot, Stable Transfection, Construct

The expression of VEGF-C, MMP-2 and E-cadherin in the stable SW480 cell lines with the overexpression and Knockdown of eIF4E A. Western blotting showed the expression of eIF4E, VEGF-C, MMP-2, and E-cadherin in the corresponding stable SW 480 cell lines with the overexpression and knockdown of eIF4E. Control, Control stable SW480 cell lines by use of lentiviral infection packaged with control empty vectors. Overexpression, the stable SW480 cell line with the overexpression of eIF4E. sheIF4E, the stable SW480 cell lines with knockdown of eIF4E. The cells were lysed for loading on SDS-PAGE. The blotting were performed by use of indicated antibodies. B. Q-PCR showed the expression of eIF4E, VEGF-C, MMP-2, and E-cadherin in the stable SW 480 cell lines with the overexpression of eIF4E. C. Q-PCR showed the expression of eIF4E, VEGF-C, MMP-2, and E-cadherin in the stable SW 480 cell lines with the knockdown of eIF4E. Control, Control stable SW480 cell lines by use of lentiviral infection packaged with control empty vectors. Overexpression of eIF4E, the stable SW480 cell line with the overexpression of eIF4E. sheIF4E, the stable SW480 cell lines with knockdown of eIF4E. *, p
Figure Legend Snippet: The expression of VEGF-C, MMP-2 and E-cadherin in the stable SW480 cell lines with the overexpression and Knockdown of eIF4E A. Western blotting showed the expression of eIF4E, VEGF-C, MMP-2, and E-cadherin in the corresponding stable SW 480 cell lines with the overexpression and knockdown of eIF4E. Control, Control stable SW480 cell lines by use of lentiviral infection packaged with control empty vectors. Overexpression, the stable SW480 cell line with the overexpression of eIF4E. sheIF4E, the stable SW480 cell lines with knockdown of eIF4E. The cells were lysed for loading on SDS-PAGE. The blotting were performed by use of indicated antibodies. B. Q-PCR showed the expression of eIF4E, VEGF-C, MMP-2, and E-cadherin in the stable SW 480 cell lines with the overexpression of eIF4E. C. Q-PCR showed the expression of eIF4E, VEGF-C, MMP-2, and E-cadherin in the stable SW 480 cell lines with the knockdown of eIF4E. Control, Control stable SW480 cell lines by use of lentiviral infection packaged with control empty vectors. Overexpression of eIF4E, the stable SW480 cell line with the overexpression of eIF4E. sheIF4E, the stable SW480 cell lines with knockdown of eIF4E. *, p

Techniques Used: Expressing, Over Expression, Western Blot, Infection, SDS Page, Polymerase Chain Reaction

The expression of eIF4E, VEGF-C, E-cadherin and MMP-2 in colon cancerous tissues A. The expression of eIF4E in normal colon tissues around cancer. B. The negative expression of eIF4E in colon cancerous tissues. C. The positive expression of eIF4E in colon cancerous tissues. D. The expression of VEGF-C in normal colon tissues around cancer. E. The negative expression of VEGF-C in colon cancerous tissues. F. The positive expression of VEGF-C in colon cancerous tissues. G. The expression of E-cadherin (E-cad) in normal colon tissues around cancer. H. The negative expression of E-cad in colon cancerous tissues. I. The positive expression of E-cad in colon cancerous tissues. J. The expression of MMP-2 in normal colon tissues around cancer. K. The negative expression of MMP-2 in colon cancerous tissues. L. The positive expression of MMP-2 in colon cancerous tissues. Bar = 50 μM.
Figure Legend Snippet: The expression of eIF4E, VEGF-C, E-cadherin and MMP-2 in colon cancerous tissues A. The expression of eIF4E in normal colon tissues around cancer. B. The negative expression of eIF4E in colon cancerous tissues. C. The positive expression of eIF4E in colon cancerous tissues. D. The expression of VEGF-C in normal colon tissues around cancer. E. The negative expression of VEGF-C in colon cancerous tissues. F. The positive expression of VEGF-C in colon cancerous tissues. G. The expression of E-cadherin (E-cad) in normal colon tissues around cancer. H. The negative expression of E-cad in colon cancerous tissues. I. The positive expression of E-cad in colon cancerous tissues. J. The expression of MMP-2 in normal colon tissues around cancer. K. The negative expression of MMP-2 in colon cancerous tissues. L. The positive expression of MMP-2 in colon cancerous tissues. Bar = 50 μM.

Techniques Used: Expressing

7) Product Images from "Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells"

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells

Journal: Acta Biochimica et Biophysica Sinica

doi: 10.1093/abbs/gmw030

ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression
Figure Legend Snippet: ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression

Techniques Used: Over Expression, Western Blot, Stable Transfection, Expressing

Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC
Figure Legend Snippet: Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC

Techniques Used: Expressing

Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly
Figure Legend Snippet: Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly

Techniques Used: Immunohistochemistry, Staining

Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells
Figure Legend Snippet: Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells

Techniques Used: Over Expression, Expressing, Activation Assay, Western Blot

Expressions of Aurora-A and MMP-2 proteins in ESCC tissue
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins in ESCC tissue

Techniques Used:

Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and

Techniques Used: Expressing, Western Blot

8) Product Images from "Expression analysis and clinical significance of eIF4E, VEGF-C, E-cadherin and MMP-2 in colorectal adenocarcinoma"

Article Title: Expression analysis and clinical significance of eIF4E, VEGF-C, E-cadherin and MMP-2 in colorectal adenocarcinoma

Journal: Oncotarget

doi: 10.18632/oncotarget.13453

RL bioluminescence from HCT-15/Rluc cells present in Lung tissue in living mice A. The HCT-15/Rluc/eIF4E cells (1.0 × 10 6 ) and corresponding HCT-15-Rluc were injected via tail-vein 3 days later. The bioluminescence seen represents the thorax region of the mouse where HCT-15/Rluc and HCT-15/Rluc/eIF4E cells are trapped in the lungs. B. The HCT-15/Rluc/VEGF-C cells (1.0 × 10 6 ) and corresponding HCT-15/Rluc were injected via tail-vein 3 days later. The bioluminescence seen represents the thorax region of the mouse where HCT-15/Rluc and HCT-15/Rluc/VEGF-C cells are trapped in the lungs. C. The HCT-15/Rluc/MMP-2 cells (1.0 × 10 6 ) and corresponding HCT-15/Rluc were injected via tail-vein 3 days later. The bioluminescence seen represents the thorax region of the mouse where HCT-15/Rluc and HCT-15/Rluc/MMP-2 cells are trapped in the lungs. D. The HCT/15-Rluc/E-cadherin cells (1.0 × 10 6 ) and corresponding HCT-15-Rluc were injected via tail-vein 3 days later. The bioluminescence seen represents the thorax region of the mouse where HCT-15/Rluc and HCT-15/Rluc/E-cadherin cells are trapped in the lungs.
Figure Legend Snippet: RL bioluminescence from HCT-15/Rluc cells present in Lung tissue in living mice A. The HCT-15/Rluc/eIF4E cells (1.0 × 10 6 ) and corresponding HCT-15-Rluc were injected via tail-vein 3 days later. The bioluminescence seen represents the thorax region of the mouse where HCT-15/Rluc and HCT-15/Rluc/eIF4E cells are trapped in the lungs. B. The HCT-15/Rluc/VEGF-C cells (1.0 × 10 6 ) and corresponding HCT-15/Rluc were injected via tail-vein 3 days later. The bioluminescence seen represents the thorax region of the mouse where HCT-15/Rluc and HCT-15/Rluc/VEGF-C cells are trapped in the lungs. C. The HCT-15/Rluc/MMP-2 cells (1.0 × 10 6 ) and corresponding HCT-15/Rluc were injected via tail-vein 3 days later. The bioluminescence seen represents the thorax region of the mouse where HCT-15/Rluc and HCT-15/Rluc/MMP-2 cells are trapped in the lungs. D. The HCT/15-Rluc/E-cadherin cells (1.0 × 10 6 ) and corresponding HCT-15-Rluc were injected via tail-vein 3 days later. The bioluminescence seen represents the thorax region of the mouse where HCT-15/Rluc and HCT-15/Rluc/E-cadherin cells are trapped in the lungs.

Techniques Used: Mouse Assay, Injection

The expression of eIF4E, VEGF-C, MMP-2 and E-cadherin in primary site of patient-derived colon cancer xenografts A. The negative expression of eIF4E in colon cancerous tissues. B. The positive expression of eIF4E in colon cancerous tissues. C. The negative expression of VEGF-C in colon cancerous tissues. D. The positive expression of VEGF-C in colon cancerous tissues. E. The negative expression of E-cadherin in colon cancerous tissues. F. The positive expression of E-cadherin in colon cancerous tissues. G. The negative expression of MMP-2 in colon cancerous tissues. H. The positive expression of MMP-2 in colon cancerous tissues. Bar = 25 μM.
Figure Legend Snippet: The expression of eIF4E, VEGF-C, MMP-2 and E-cadherin in primary site of patient-derived colon cancer xenografts A. The negative expression of eIF4E in colon cancerous tissues. B. The positive expression of eIF4E in colon cancerous tissues. C. The negative expression of VEGF-C in colon cancerous tissues. D. The positive expression of VEGF-C in colon cancerous tissues. E. The negative expression of E-cadherin in colon cancerous tissues. F. The positive expression of E-cadherin in colon cancerous tissues. G. The negative expression of MMP-2 in colon cancerous tissues. H. The positive expression of MMP-2 in colon cancerous tissues. Bar = 25 μM.

Techniques Used: Expressing, Derivative Assay

The growth rates of colon cancer cell lines HCT-15/Rluc with overexpression of E-cadherin, MMP-2, VEGF-c and eIF4E and parental HCT-15/Rluc A. Western blotting showed that the stable cell lines including HCT-15/Rluc/E-cadherin, HCT-15/Rluc/VEGF-C, HCT-15/Rluc/MMP-2, and HCT-15/Rluc/eIF4E were constructed successfully. B. The stable colon cancer cell lines with overexpression of E-cadherin, MMP-2, VEGF-c and eIF4E as well as parental HCT-15/Rluc cell line showed different growth rates.
Figure Legend Snippet: The growth rates of colon cancer cell lines HCT-15/Rluc with overexpression of E-cadherin, MMP-2, VEGF-c and eIF4E and parental HCT-15/Rluc A. Western blotting showed that the stable cell lines including HCT-15/Rluc/E-cadherin, HCT-15/Rluc/VEGF-C, HCT-15/Rluc/MMP-2, and HCT-15/Rluc/eIF4E were constructed successfully. B. The stable colon cancer cell lines with overexpression of E-cadherin, MMP-2, VEGF-c and eIF4E as well as parental HCT-15/Rluc cell line showed different growth rates.

Techniques Used: Over Expression, Western Blot, Stable Transfection, Construct

The expression of VEGF-C, MMP-2 and E-cadherin in the stable SW480 cell lines with the overexpression and Knockdown of eIF4E A. Western blotting showed the expression of eIF4E, VEGF-C, MMP-2, and E-cadherin in the corresponding stable SW 480 cell lines with the overexpression and knockdown of eIF4E. Control, Control stable SW480 cell lines by use of lentiviral infection packaged with control empty vectors. Overexpression, the stable SW480 cell line with the overexpression of eIF4E. sheIF4E, the stable SW480 cell lines with knockdown of eIF4E. The cells were lysed for loading on SDS-PAGE. The blotting were performed by use of indicated antibodies. B. Q-PCR showed the expression of eIF4E, VEGF-C, MMP-2, and E-cadherin in the stable SW 480 cell lines with the overexpression of eIF4E. C. Q-PCR showed the expression of eIF4E, VEGF-C, MMP-2, and E-cadherin in the stable SW 480 cell lines with the knockdown of eIF4E. Control, Control stable SW480 cell lines by use of lentiviral infection packaged with control empty vectors. Overexpression of eIF4E, the stable SW480 cell line with the overexpression of eIF4E. sheIF4E, the stable SW480 cell lines with knockdown of eIF4E. *, p
Figure Legend Snippet: The expression of VEGF-C, MMP-2 and E-cadherin in the stable SW480 cell lines with the overexpression and Knockdown of eIF4E A. Western blotting showed the expression of eIF4E, VEGF-C, MMP-2, and E-cadherin in the corresponding stable SW 480 cell lines with the overexpression and knockdown of eIF4E. Control, Control stable SW480 cell lines by use of lentiviral infection packaged with control empty vectors. Overexpression, the stable SW480 cell line with the overexpression of eIF4E. sheIF4E, the stable SW480 cell lines with knockdown of eIF4E. The cells were lysed for loading on SDS-PAGE. The blotting were performed by use of indicated antibodies. B. Q-PCR showed the expression of eIF4E, VEGF-C, MMP-2, and E-cadherin in the stable SW 480 cell lines with the overexpression of eIF4E. C. Q-PCR showed the expression of eIF4E, VEGF-C, MMP-2, and E-cadherin in the stable SW 480 cell lines with the knockdown of eIF4E. Control, Control stable SW480 cell lines by use of lentiviral infection packaged with control empty vectors. Overexpression of eIF4E, the stable SW480 cell line with the overexpression of eIF4E. sheIF4E, the stable SW480 cell lines with knockdown of eIF4E. *, p

Techniques Used: Expressing, Over Expression, Western Blot, Infection, SDS Page, Polymerase Chain Reaction

The expression of eIF4E, VEGF-C, E-cadherin and MMP-2 in colon cancerous tissues A. The expression of eIF4E in normal colon tissues around cancer. B. The negative expression of eIF4E in colon cancerous tissues. C. The positive expression of eIF4E in colon cancerous tissues. D. The expression of VEGF-C in normal colon tissues around cancer. E. The negative expression of VEGF-C in colon cancerous tissues. F. The positive expression of VEGF-C in colon cancerous tissues. G. The expression of E-cadherin (E-cad) in normal colon tissues around cancer. H. The negative expression of E-cad in colon cancerous tissues. I. The positive expression of E-cad in colon cancerous tissues. J. The expression of MMP-2 in normal colon tissues around cancer. K. The negative expression of MMP-2 in colon cancerous tissues. L. The positive expression of MMP-2 in colon cancerous tissues. Bar = 50 μM.
Figure Legend Snippet: The expression of eIF4E, VEGF-C, E-cadherin and MMP-2 in colon cancerous tissues A. The expression of eIF4E in normal colon tissues around cancer. B. The negative expression of eIF4E in colon cancerous tissues. C. The positive expression of eIF4E in colon cancerous tissues. D. The expression of VEGF-C in normal colon tissues around cancer. E. The negative expression of VEGF-C in colon cancerous tissues. F. The positive expression of VEGF-C in colon cancerous tissues. G. The expression of E-cadherin (E-cad) in normal colon tissues around cancer. H. The negative expression of E-cad in colon cancerous tissues. I. The positive expression of E-cad in colon cancerous tissues. J. The expression of MMP-2 in normal colon tissues around cancer. K. The negative expression of MMP-2 in colon cancerous tissues. L. The positive expression of MMP-2 in colon cancerous tissues. Bar = 50 μM.

Techniques Used: Expressing

9) Product Images from "Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells"

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells

Journal: Acta Biochimica et Biophysica Sinica

doi: 10.1093/abbs/gmw030

ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression
Figure Legend Snippet: ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression

Techniques Used: Over Expression, Western Blot, Stable Transfection, Expressing

Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC
Figure Legend Snippet: Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC

Techniques Used: Expressing

Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly
Figure Legend Snippet: Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly

Techniques Used: Immunohistochemistry, Staining

Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells
Figure Legend Snippet: Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells

Techniques Used: Over Expression, Expressing, Activation Assay, Western Blot

Expressions of Aurora-A and MMP-2 proteins in ESCC tissue
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins in ESCC tissue

Techniques Used:

Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and

Techniques Used: Expressing, Western Blot

10) Product Images from "Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells"

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells

Journal: Acta Biochimica et Biophysica Sinica

doi: 10.1093/abbs/gmw030

ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression
Figure Legend Snippet: ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression

Techniques Used: Over Expression, Western Blot, Stable Transfection, Expressing

Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC
Figure Legend Snippet: Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC

Techniques Used: Expressing

Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly
Figure Legend Snippet: Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly

Techniques Used: Immunohistochemistry, Staining

Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells
Figure Legend Snippet: Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells

Techniques Used: Over Expression, Expressing, Activation Assay, Western Blot

Expressions of Aurora-A and MMP-2 proteins in ESCC tissue
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins in ESCC tissue

Techniques Used:

Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and

Techniques Used: Expressing, Western Blot

11) Product Images from "Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells"

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells

Journal: Acta Biochimica et Biophysica Sinica

doi: 10.1093/abbs/gmw030

ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression
Figure Legend Snippet: ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression

Techniques Used: Over Expression, Western Blot, Stable Transfection, Expressing

Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC
Figure Legend Snippet: Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC

Techniques Used: Expressing

Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly
Figure Legend Snippet: Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly

Techniques Used: Immunohistochemistry, Staining

Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells
Figure Legend Snippet: Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells

Techniques Used: Over Expression, Expressing, Activation Assay, Western Blot

Expressions of Aurora-A and MMP-2 proteins in ESCC tissue
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins in ESCC tissue

Techniques Used:

Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and

Techniques Used: Expressing, Western Blot

12) Product Images from "Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells"

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells

Journal: Acta Biochimica et Biophysica Sinica

doi: 10.1093/abbs/gmw030

ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression
Figure Legend Snippet: ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression

Techniques Used: Over Expression, Western Blot, Stable Transfection, Expressing

Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC
Figure Legend Snippet: Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC

Techniques Used: Expressing

Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly
Figure Legend Snippet: Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly

Techniques Used: Immunohistochemistry, Staining

Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells
Figure Legend Snippet: Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells

Techniques Used: Over Expression, Expressing, Activation Assay, Western Blot

Expressions of Aurora-A and MMP-2 proteins in ESCC tissue
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins in ESCC tissue

Techniques Used:

Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and

Techniques Used: Expressing, Western Blot

13) Product Images from "Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells"

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells

Journal: Acta Biochimica et Biophysica Sinica

doi: 10.1093/abbs/gmw030

ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression
Figure Legend Snippet: ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression

Techniques Used: Over Expression, Western Blot, Stable Transfection, Expressing

Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC
Figure Legend Snippet: Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC

Techniques Used: Expressing

Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly
Figure Legend Snippet: Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly

Techniques Used: Immunohistochemistry, Staining

Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells
Figure Legend Snippet: Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells

Techniques Used: Over Expression, Expressing, Activation Assay, Western Blot

Expressions of Aurora-A and MMP-2 proteins in ESCC tissue
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins in ESCC tissue

Techniques Used:

Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and

Techniques Used: Expressing, Western Blot

14) Product Images from "Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells"

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells

Journal: Acta Biochimica et Biophysica Sinica

doi: 10.1093/abbs/gmw030

ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression
Figure Legend Snippet: ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression

Techniques Used: Over Expression, Western Blot, Stable Transfection, Expressing

Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC
Figure Legend Snippet: Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC

Techniques Used: Expressing

Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly
Figure Legend Snippet: Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly

Techniques Used: Immunohistochemistry, Staining

Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells
Figure Legend Snippet: Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells

Techniques Used: Over Expression, Expressing, Activation Assay, Western Blot

Expressions of Aurora-A and MMP-2 proteins in ESCC tissue
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins in ESCC tissue

Techniques Used:

Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and

Techniques Used: Expressing, Western Blot

15) Product Images from "Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells"

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells

Journal: Acta Biochimica et Biophysica Sinica

doi: 10.1093/abbs/gmw030

ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression
Figure Legend Snippet: ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression

Techniques Used: Over Expression, Western Blot, Stable Transfection, Expressing

Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC
Figure Legend Snippet: Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC

Techniques Used: Expressing

Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly
Figure Legend Snippet: Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly

Techniques Used: Immunohistochemistry, Staining

Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells
Figure Legend Snippet: Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells

Techniques Used: Over Expression, Expressing, Activation Assay, Western Blot

Expressions of Aurora-A and MMP-2 proteins in ESCC tissue
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins in ESCC tissue

Techniques Used:

Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and

Techniques Used: Expressing, Western Blot

16) Product Images from "Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells"

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells

Journal: Acta Biochimica et Biophysica Sinica

doi: 10.1093/abbs/gmw030

ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression
Figure Legend Snippet: ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression

Techniques Used: Over Expression, Western Blot, Stable Transfection, Expressing

Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC
Figure Legend Snippet: Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC

Techniques Used: Expressing

Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly
Figure Legend Snippet: Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly

Techniques Used: Immunohistochemistry, Staining

Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells
Figure Legend Snippet: Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells

Techniques Used: Over Expression, Expressing, Activation Assay, Western Blot

Expressions of Aurora-A and MMP-2 proteins in ESCC tissue
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins in ESCC tissue

Techniques Used:

Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and

Techniques Used: Expressing, Western Blot

17) Product Images from "Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells"

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells

Journal: Acta Biochimica et Biophysica Sinica

doi: 10.1093/abbs/gmw030

ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression
Figure Legend Snippet: ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression

Techniques Used: Over Expression, Western Blot, Stable Transfection, Expressing

Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC
Figure Legend Snippet: Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC

Techniques Used: Expressing

Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly
Figure Legend Snippet: Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly

Techniques Used: Immunohistochemistry, Staining

Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells
Figure Legend Snippet: Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells

Techniques Used: Over Expression, Expressing, Activation Assay, Western Blot

Expressions of Aurora-A and MMP-2 proteins in ESCC tissue
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins in ESCC tissue

Techniques Used:

Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and

Techniques Used: Expressing, Western Blot

18) Product Images from "Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells"

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells

Journal: Acta Biochimica et Biophysica Sinica

doi: 10.1093/abbs/gmw030

ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression
Figure Legend Snippet: ESCC cell invasion is promoted by Aurora-A overexpression and attenuated by the MMP-2 inhibitor I (A) The levels of fusion protein GFP-Aurora-A and MMP-2 were detected by western blot analysis after stable transfection of GFP-Aurora-A expression

Techniques Used: Over Expression, Western Blot, Stable Transfection, Expressing

Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC
Figure Legend Snippet: Expression of Aurora-A is positively correlated with expression of MMP - 2 in ESCC

Techniques Used: Expressing

Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly
Figure Legend Snippet: Representative immunohistochemical stainings of MMP-2 in ESCC and normal adjacent tissues on TMA (A) In normal adjacent esophageal tissues, MMP-2 showed negative cytoplasmic and weak nuclear staining. In ESCC tissues, MMP-2 staining was weakly

Techniques Used: Immunohistochemistry, Staining

Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells
Figure Legend Snippet: Aurora-A overexpression upregulates MMP-2 expression via AKT/NF-κB activation in ESCC cells (A) The nuclear protein level of NF-κB p65 was detected by western blot analysis in Aurora-A-overexpressing and control cells. (B) Cells

Techniques Used: Over Expression, Expressing, Activation Assay, Western Blot

Expressions of Aurora-A and MMP-2 proteins in ESCC tissue
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins in ESCC tissue

Techniques Used:

Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and
Figure Legend Snippet: Expressions of Aurora-A and MMP-2 proteins as well as cell invasion capability in ESCC cell lines (A) The expression levels of Aurora-A and MMP-2 were detected by western blot analysis in KYSE150 and EC9706 cells. (B) The relative Aurora-A and

Techniques Used: Expressing, Western Blot

19) Product Images from "Thrombin-dependent MMP-2 Activity Is Regulated by Heparan Sulfate *"

Article Title: Thrombin-dependent MMP-2 Activity Is Regulated by Heparan Sulfate *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M110.171595

Heparan sulfate proteoglycans are required for thrombin-mediated activation of pro-MMP-2. A , Western blot analysis using an anti-MMP-2 antibody of 20 μg/ml pro-MMP-was 2 incubated with 50 n m thrombin in HBMEC cells for the indicated times. Cells
Figure Legend Snippet: Heparan sulfate proteoglycans are required for thrombin-mediated activation of pro-MMP-2. A , Western blot analysis using an anti-MMP-2 antibody of 20 μg/ml pro-MMP-was 2 incubated with 50 n m thrombin in HBMEC cells for the indicated times. Cells

Techniques Used: Activation Assay, Western Blot, Incubation

Binding of heparan sulfate to thrombin elicits a decrease in thrombin-mediated MMP-2 degradation and a concomitant increase in activation. A , shown is the effect of heparan sulfate on the activation and degradation of MMP-2 and fibrinogen cleavage by
Figure Legend Snippet: Binding of heparan sulfate to thrombin elicits a decrease in thrombin-mediated MMP-2 degradation and a concomitant increase in activation. A , shown is the effect of heparan sulfate on the activation and degradation of MMP-2 and fibrinogen cleavage by

Techniques Used: Binding Assay, Activation Assay

Simultaneous binding of MMP-2 to exosite 1 and 2 is essential for its degradation by thrombin. A , Western blot ( IB ) analysis using an anti-myc antibody of 20 μg/ml pro-MMP-2 incubated with 50 n m thrombin in the presence of heparan sulfate (40
Figure Legend Snippet: Simultaneous binding of MMP-2 to exosite 1 and 2 is essential for its degradation by thrombin. A , Western blot ( IB ) analysis using an anti-myc antibody of 20 μg/ml pro-MMP-2 incubated with 50 n m thrombin in the presence of heparan sulfate (40

Techniques Used: Binding Assay, Western Blot, Incubation

Expression of syndecan-1 increases thrombin-mediated activation of pro-MMP-2 in K562 cells. A , flow cytometric analysis shows cell surface expression of heparan sulfate in K562 cells stably transfected with syndecan-1 is shown. A sample lacking primary
Figure Legend Snippet: Expression of syndecan-1 increases thrombin-mediated activation of pro-MMP-2 in K562 cells. A , flow cytometric analysis shows cell surface expression of heparan sulfate in K562 cells stably transfected with syndecan-1 is shown. A sample lacking primary

Techniques Used: Expressing, Activation Assay, Flow Cytometry, Stable Transfection, Transfection

Degradation of MMP-2 by thrombin is dependent on the hemopexin-like domain. A , Western blot ( IB ) analysis using an anti-myc antibody of 20 μg/ml pro-MMP-2 incubated with 50 n m thrombin in the presence of recombinant hemopexin-like domain of MMP-2
Figure Legend Snippet: Degradation of MMP-2 by thrombin is dependent on the hemopexin-like domain. A , Western blot ( IB ) analysis using an anti-myc antibody of 20 μg/ml pro-MMP-2 incubated with 50 n m thrombin in the presence of recombinant hemopexin-like domain of MMP-2

Techniques Used: Western Blot, Incubation, Recombinant

Heparan sulfate increases thrombin-mediated activation of pro-MMP-2 under cell-free conditions. A , Western blotting using an anti-MMP-2 antibody of 20 μg/ml pro-MMP-2 incubated with 50 n m thrombin in the presence of heparan sulfate under cell-free
Figure Legend Snippet: Heparan sulfate increases thrombin-mediated activation of pro-MMP-2 under cell-free conditions. A , Western blotting using an anti-MMP-2 antibody of 20 μg/ml pro-MMP-2 incubated with 50 n m thrombin in the presence of heparan sulfate under cell-free

Techniques Used: Activation Assay, Western Blot, Incubation

20) Product Images from "CXCR3 activation by lentivirus infection suppresses neuronal autophagy: neuroprotective effects of antiretroviral therapy"

Article Title: CXCR3 activation by lentivirus infection suppresses neuronal autophagy: neuroprotective effects of antiretroviral therapy

Journal: The FASEB Journal

doi: 10.1096/fj.08-128819

Brain expression of CXCL12, MMP-2, and CXCR3 in FIV infection. A ) Immunoblotting showed increased full-length (1-67) and cleaved (5-67) CXCL12 in brains of FIV − compared with mock-infected animals. B ) CXCL12, CXCL12(5-67), MMP-2, and CXCR3 immunoreactivities were increased in brains of FIV + animals but were suppressed by concurrent ddI treatment. C ) MMP-2 immunoreactivity (IR) showed minimal induction in FIV + animals with and without ddI treatment. D , E ) CXCL12(1-67) ( D ) and CXCL12(5-67) ( E ) were induced by FIV infection, but these changes were reversed by ddI treatment. F ) CXCR3 showed minimal induction of both immunoreactive bands in FIV infection and was not affected by ddI treatment. * P
Figure Legend Snippet: Brain expression of CXCL12, MMP-2, and CXCR3 in FIV infection. A ) Immunoblotting showed increased full-length (1-67) and cleaved (5-67) CXCL12 in brains of FIV − compared with mock-infected animals. B ) CXCL12, CXCL12(5-67), MMP-2, and CXCR3 immunoreactivities were increased in brains of FIV + animals but were suppressed by concurrent ddI treatment. C ) MMP-2 immunoreactivity (IR) showed minimal induction in FIV + animals with and without ddI treatment. D , E ) CXCL12(1-67) ( D ) and CXCL12(5-67) ( E ) were induced by FIV infection, but these changes were reversed by ddI treatment. F ) CXCR3 showed minimal induction of both immunoreactive bands in FIV infection and was not affected by ddI treatment. * P

Techniques Used: Expressing, Infection

Neurobehavioral performance, viral burden, and neuroinflammation in FIV infection. A ) FIV-infected animals (FIV+) exhibit greater neurobehavioral deficits than mock-infected controls (FIV−), while ddI treatment reversed neurobehavioral deficits in FIV-infected animals (FIV+/ddI) and had no effects on mock-infected animals. B ) ddI treatment suppressed viral load in plasma of FIV-infected animals. C ) Glycoprotein, F4/80 , expressed in activated myeloid cells showed increased transcript abundance in FIV-infected animals, which was reversed by ddI treatment. D ) CXCL12 transcript levels were higher in FIV-infected animals but were not reduced by ddI treatment. E ) MMP-2 transcripts were unregulated in FIV-infected animals but were suppressed with ddI treatment. F ) GFAP transcriptional activity was not affected by FIV infection or ddI treatment. * P
Figure Legend Snippet: Neurobehavioral performance, viral burden, and neuroinflammation in FIV infection. A ) FIV-infected animals (FIV+) exhibit greater neurobehavioral deficits than mock-infected controls (FIV−), while ddI treatment reversed neurobehavioral deficits in FIV-infected animals (FIV+/ddI) and had no effects on mock-infected animals. B ) ddI treatment suppressed viral load in plasma of FIV-infected animals. C ) Glycoprotein, F4/80 , expressed in activated myeloid cells showed increased transcript abundance in FIV-infected animals, which was reversed by ddI treatment. D ) CXCL12 transcript levels were higher in FIV-infected animals but were not reduced by ddI treatment. E ) MMP-2 transcripts were unregulated in FIV-infected animals but were suppressed with ddI treatment. F ) GFAP transcriptional activity was not affected by FIV infection or ddI treatment. * P

Techniques Used: Infection, Activity Assay

Suppression of neuronal autophagy and its regulation by antiretroviral therapy. ddI suppressed viral burden in the peripheral circulation, diminishing leukocyte activation and CNS entry with an ensuing reduction in MMP-2 production and its actions, thereby preventing the induction of CXCL12(5-12) with its adverse effects on neuronal autophagy and survival.
Figure Legend Snippet: Suppression of neuronal autophagy and its regulation by antiretroviral therapy. ddI suppressed viral burden in the peripheral circulation, diminishing leukocyte activation and CNS entry with an ensuing reduction in MMP-2 production and its actions, thereby preventing the induction of CXCL12(5-12) with its adverse effects on neuronal autophagy and survival.

Techniques Used: Activation Assay

21) Product Images from "CXCR3 activation by lentivirus infection suppresses neuronal autophagy: neuroprotective effects of antiretroviral therapy"

Article Title: CXCR3 activation by lentivirus infection suppresses neuronal autophagy: neuroprotective effects of antiretroviral therapy

Journal: The FASEB Journal

doi: 10.1096/fj.08-128819

Brain expression of CXCL12, MMP-2, and CXCR3 in FIV infection. A ) Immunoblotting showed increased full-length (1-67) and cleaved (5-67) CXCL12 in brains of FIV − compared with mock-infected animals. B ) CXCL12, CXCL12(5-67), MMP-2, and CXCR3 immunoreactivities were increased in brains of FIV + animals but were suppressed by concurrent ddI treatment. C ) MMP-2 immunoreactivity (IR) showed minimal induction in FIV + animals with and without ddI treatment. D , E ) CXCL12(1-67) ( D ) and CXCL12(5-67) ( E ) were induced by FIV infection, but these changes were reversed by ddI treatment. F ) CXCR3 showed minimal induction of both immunoreactive bands in FIV infection and was not affected by ddI treatment. * P
Figure Legend Snippet: Brain expression of CXCL12, MMP-2, and CXCR3 in FIV infection. A ) Immunoblotting showed increased full-length (1-67) and cleaved (5-67) CXCL12 in brains of FIV − compared with mock-infected animals. B ) CXCL12, CXCL12(5-67), MMP-2, and CXCR3 immunoreactivities were increased in brains of FIV + animals but were suppressed by concurrent ddI treatment. C ) MMP-2 immunoreactivity (IR) showed minimal induction in FIV + animals with and without ddI treatment. D , E ) CXCL12(1-67) ( D ) and CXCL12(5-67) ( E ) were induced by FIV infection, but these changes were reversed by ddI treatment. F ) CXCR3 showed minimal induction of both immunoreactive bands in FIV infection and was not affected by ddI treatment. * P

Techniques Used: Expressing, Infection

Neurobehavioral performance, viral burden, and neuroinflammation in FIV infection. A ) FIV-infected animals (FIV+) exhibit greater neurobehavioral deficits than mock-infected controls (FIV−), while ddI treatment reversed neurobehavioral deficits in FIV-infected animals (FIV+/ddI) and had no effects on mock-infected animals. B ) ddI treatment suppressed viral load in plasma of FIV-infected animals. C ) Glycoprotein, F4/80 , expressed in activated myeloid cells showed increased transcript abundance in FIV-infected animals, which was reversed by ddI treatment. D ) CXCL12 transcript levels were higher in FIV-infected animals but were not reduced by ddI treatment. E ) MMP-2 transcripts were unregulated in FIV-infected animals but were suppressed with ddI treatment. F ) GFAP transcriptional activity was not affected by FIV infection or ddI treatment. * P
Figure Legend Snippet: Neurobehavioral performance, viral burden, and neuroinflammation in FIV infection. A ) FIV-infected animals (FIV+) exhibit greater neurobehavioral deficits than mock-infected controls (FIV−), while ddI treatment reversed neurobehavioral deficits in FIV-infected animals (FIV+/ddI) and had no effects on mock-infected animals. B ) ddI treatment suppressed viral load in plasma of FIV-infected animals. C ) Glycoprotein, F4/80 , expressed in activated myeloid cells showed increased transcript abundance in FIV-infected animals, which was reversed by ddI treatment. D ) CXCL12 transcript levels were higher in FIV-infected animals but were not reduced by ddI treatment. E ) MMP-2 transcripts were unregulated in FIV-infected animals but were suppressed with ddI treatment. F ) GFAP transcriptional activity was not affected by FIV infection or ddI treatment. * P

Techniques Used: Infection, Activity Assay

Suppression of neuronal autophagy and its regulation by antiretroviral therapy. ddI suppressed viral burden in the peripheral circulation, diminishing leukocyte activation and CNS entry with an ensuing reduction in MMP-2 production and its actions, thereby preventing the induction of CXCL12(5-12) with its adverse effects on neuronal autophagy and survival.
Figure Legend Snippet: Suppression of neuronal autophagy and its regulation by antiretroviral therapy. ddI suppressed viral burden in the peripheral circulation, diminishing leukocyte activation and CNS entry with an ensuing reduction in MMP-2 production and its actions, thereby preventing the induction of CXCL12(5-12) with its adverse effects on neuronal autophagy and survival.

Techniques Used: Activation Assay

22) Product Images from "CXCR3 activation by lentivirus infection suppresses neuronal autophagy: neuroprotective effects of antiretroviral therapy"

Article Title: CXCR3 activation by lentivirus infection suppresses neuronal autophagy: neuroprotective effects of antiretroviral therapy

Journal: The FASEB Journal

doi: 10.1096/fj.08-128819

Brain expression of CXCL12, MMP-2, and CXCR3 in FIV infection. A ) Immunoblotting showed increased full-length (1-67) and cleaved (5-67) CXCL12 in brains of FIV − compared with mock-infected animals. B ) CXCL12, CXCL12(5-67), MMP-2, and CXCR3 immunoreactivities were increased in brains of FIV + animals but were suppressed by concurrent ddI treatment. C ) MMP-2 immunoreactivity (IR) showed minimal induction in FIV + animals with and without ddI treatment. D , E ) CXCL12(1-67) ( D ) and CXCL12(5-67) ( E ) were induced by FIV infection, but these changes were reversed by ddI treatment. F ) CXCR3 showed minimal induction of both immunoreactive bands in FIV infection and was not affected by ddI treatment. * P
Figure Legend Snippet: Brain expression of CXCL12, MMP-2, and CXCR3 in FIV infection. A ) Immunoblotting showed increased full-length (1-67) and cleaved (5-67) CXCL12 in brains of FIV − compared with mock-infected animals. B ) CXCL12, CXCL12(5-67), MMP-2, and CXCR3 immunoreactivities were increased in brains of FIV + animals but were suppressed by concurrent ddI treatment. C ) MMP-2 immunoreactivity (IR) showed minimal induction in FIV + animals with and without ddI treatment. D , E ) CXCL12(1-67) ( D ) and CXCL12(5-67) ( E ) were induced by FIV infection, but these changes were reversed by ddI treatment. F ) CXCR3 showed minimal induction of both immunoreactive bands in FIV infection and was not affected by ddI treatment. * P

Techniques Used: Expressing, Infection

Neurobehavioral performance, viral burden, and neuroinflammation in FIV infection. A ) FIV-infected animals (FIV+) exhibit greater neurobehavioral deficits than mock-infected controls (FIV−), while ddI treatment reversed neurobehavioral deficits in FIV-infected animals (FIV+/ddI) and had no effects on mock-infected animals. B ) ddI treatment suppressed viral load in plasma of FIV-infected animals. C ) Glycoprotein, F4/80 , expressed in activated myeloid cells showed increased transcript abundance in FIV-infected animals, which was reversed by ddI treatment. D ) CXCL12 transcript levels were higher in FIV-infected animals but were not reduced by ddI treatment. E ) MMP-2 transcripts were unregulated in FIV-infected animals but were suppressed with ddI treatment. F ) GFAP transcriptional activity was not affected by FIV infection or ddI treatment. * P
Figure Legend Snippet: Neurobehavioral performance, viral burden, and neuroinflammation in FIV infection. A ) FIV-infected animals (FIV+) exhibit greater neurobehavioral deficits than mock-infected controls (FIV−), while ddI treatment reversed neurobehavioral deficits in FIV-infected animals (FIV+/ddI) and had no effects on mock-infected animals. B ) ddI treatment suppressed viral load in plasma of FIV-infected animals. C ) Glycoprotein, F4/80 , expressed in activated myeloid cells showed increased transcript abundance in FIV-infected animals, which was reversed by ddI treatment. D ) CXCL12 transcript levels were higher in FIV-infected animals but were not reduced by ddI treatment. E ) MMP-2 transcripts were unregulated in FIV-infected animals but were suppressed with ddI treatment. F ) GFAP transcriptional activity was not affected by FIV infection or ddI treatment. * P

Techniques Used: Infection, Activity Assay

Suppression of neuronal autophagy and its regulation by antiretroviral therapy. ddI suppressed viral burden in the peripheral circulation, diminishing leukocyte activation and CNS entry with an ensuing reduction in MMP-2 production and its actions, thereby preventing the induction of CXCL12(5-12) with its adverse effects on neuronal autophagy and survival.
Figure Legend Snippet: Suppression of neuronal autophagy and its regulation by antiretroviral therapy. ddI suppressed viral burden in the peripheral circulation, diminishing leukocyte activation and CNS entry with an ensuing reduction in MMP-2 production and its actions, thereby preventing the induction of CXCL12(5-12) with its adverse effects on neuronal autophagy and survival.

Techniques Used: Activation Assay

23) Product Images from "CXCR3 activation by lentivirus infection suppresses neuronal autophagy: neuroprotective effects of antiretroviral therapy"

Article Title: CXCR3 activation by lentivirus infection suppresses neuronal autophagy: neuroprotective effects of antiretroviral therapy

Journal: The FASEB Journal

doi: 10.1096/fj.08-128819

Brain expression of CXCL12, MMP-2, and CXCR3 in FIV infection. A ) Immunoblotting showed increased full-length (1-67) and cleaved (5-67) CXCL12 in brains of FIV − compared with mock-infected animals. B ) CXCL12, CXCL12(5-67), MMP-2, and CXCR3 immunoreactivities were increased in brains of FIV + animals but were suppressed by concurrent ddI treatment. C ) MMP-2 immunoreactivity (IR) showed minimal induction in FIV + animals with and without ddI treatment. D , E ) CXCL12(1-67) ( D ) and CXCL12(5-67) ( E ) were induced by FIV infection, but these changes were reversed by ddI treatment. F ) CXCR3 showed minimal induction of both immunoreactive bands in FIV infection and was not affected by ddI treatment. * P
Figure Legend Snippet: Brain expression of CXCL12, MMP-2, and CXCR3 in FIV infection. A ) Immunoblotting showed increased full-length (1-67) and cleaved (5-67) CXCL12 in brains of FIV − compared with mock-infected animals. B ) CXCL12, CXCL12(5-67), MMP-2, and CXCR3 immunoreactivities were increased in brains of FIV + animals but were suppressed by concurrent ddI treatment. C ) MMP-2 immunoreactivity (IR) showed minimal induction in FIV + animals with and without ddI treatment. D , E ) CXCL12(1-67) ( D ) and CXCL12(5-67) ( E ) were induced by FIV infection, but these changes were reversed by ddI treatment. F ) CXCR3 showed minimal induction of both immunoreactive bands in FIV infection and was not affected by ddI treatment. * P

Techniques Used: Expressing, Infection

Neurobehavioral performance, viral burden, and neuroinflammation in FIV infection. A ) FIV-infected animals (FIV+) exhibit greater neurobehavioral deficits than mock-infected controls (FIV−), while ddI treatment reversed neurobehavioral deficits in FIV-infected animals (FIV+/ddI) and had no effects on mock-infected animals. B ) ddI treatment suppressed viral load in plasma of FIV-infected animals. C ) Glycoprotein, F4/80 , expressed in activated myeloid cells showed increased transcript abundance in FIV-infected animals, which was reversed by ddI treatment. D ) CXCL12 transcript levels were higher in FIV-infected animals but were not reduced by ddI treatment. E ) MMP-2 transcripts were unregulated in FIV-infected animals but were suppressed with ddI treatment. F ) GFAP transcriptional activity was not affected by FIV infection or ddI treatment. * P
Figure Legend Snippet: Neurobehavioral performance, viral burden, and neuroinflammation in FIV infection. A ) FIV-infected animals (FIV+) exhibit greater neurobehavioral deficits than mock-infected controls (FIV−), while ddI treatment reversed neurobehavioral deficits in FIV-infected animals (FIV+/ddI) and had no effects on mock-infected animals. B ) ddI treatment suppressed viral load in plasma of FIV-infected animals. C ) Glycoprotein, F4/80 , expressed in activated myeloid cells showed increased transcript abundance in FIV-infected animals, which was reversed by ddI treatment. D ) CXCL12 transcript levels were higher in FIV-infected animals but were not reduced by ddI treatment. E ) MMP-2 transcripts were unregulated in FIV-infected animals but were suppressed with ddI treatment. F ) GFAP transcriptional activity was not affected by FIV infection or ddI treatment. * P

Techniques Used: Infection, Activity Assay

Suppression of neuronal autophagy and its regulation by antiretroviral therapy. ddI suppressed viral burden in the peripheral circulation, diminishing leukocyte activation and CNS entry with an ensuing reduction in MMP-2 production and its actions, thereby preventing the induction of CXCL12(5-12) with its adverse effects on neuronal autophagy and survival.
Figure Legend Snippet: Suppression of neuronal autophagy and its regulation by antiretroviral therapy. ddI suppressed viral burden in the peripheral circulation, diminishing leukocyte activation and CNS entry with an ensuing reduction in MMP-2 production and its actions, thereby preventing the induction of CXCL12(5-12) with its adverse effects on neuronal autophagy and survival.

Techniques Used: Activation Assay

24) Product Images from "MMP‐2 and MMP‐13 affect vasculogenic mimicry formation in large cell lung cancer"

Article Title: MMP‐2 and MMP‐13 affect vasculogenic mimicry formation in large cell lung cancer

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.13283

MMP ‐13 and  MMP ‐2 expression in different tumour stages. ( A ) Tumour volume at different weeks. ( B ) Endomucin/ PAS  double‐staining in H460 xenografts showed decreased  VM  but increased  EDV  numbers (Bar = 100 μm). The black arrow shows  VM  formation. The red arrow shows the  EDV s. ( C ) The expression levels of  MMP ‐13 showed a gradually increasing trend with tumour growth. Meanwhile, no significant difference in the  MMP ‐2 expression was observed at different tumour growth stages (Bar = 100 μm).  WB  showed  MMP ‐13 gradually increasing, but Ln‐5 and  EGFR  showed a decreasing trend with tumour growth.  MMP ‐2 expression was observed at different tumour growth stages.
Figure Legend Snippet: MMP ‐13 and MMP ‐2 expression in different tumour stages. ( A ) Tumour volume at different weeks. ( B ) Endomucin/ PAS double‐staining in H460 xenografts showed decreased VM but increased EDV numbers (Bar = 100 μm). The black arrow shows VM formation. The red arrow shows the EDV s. ( C ) The expression levels of MMP ‐13 showed a gradually increasing trend with tumour growth. Meanwhile, no significant difference in the MMP ‐2 expression was observed at different tumour growth stages (Bar = 100 μm). WB showed MMP ‐13 gradually increasing, but Ln‐5 and EGFR showed a decreasing trend with tumour growth. MMP ‐2 expression was observed at different tumour growth stages.

Techniques Used: Expressing, Double Staining, Western Blot

Cleavage of Ln‐5 by  MMP ‐13 was detected with a silver staining kit and  LC ‐ MS / MS . ( A ) Human Ln‐5 was cleaved by  MMP ‐2 and  MMP ‐13 at 37°C for 6 hrs, and the cleavage fragments were separated on a 12%  SDS ‐ PAGE  gel and detected with a silver staining kit. The left lane shows the 140‐ and 100‐ kD a forms of the intact Ln‐5γ2 chain.  MMP ‐2 generated an 80 kD Ln‐5γ2x fragment and a very faint 66 kD fragment.  MMP ‐13 further generated low molecular weight fragments that were approximately 20 kD. ( B )  LC ‐ MS / MS  detection showed that the 20 kD fragment generated by  MMP ‐13 was part of the Ln‐5γ2 chain. ( C )  MMP ‐13 cleaved the Ln‐5γ2 chain at Lys‐Cys (540–541) and Arg‐Leu (694–695). This fragment belongs to the Ln‐5γ2 chain  III  and IV domain.
Figure Legend Snippet: Cleavage of Ln‐5 by MMP ‐13 was detected with a silver staining kit and LC ‐ MS / MS . ( A ) Human Ln‐5 was cleaved by MMP ‐2 and MMP ‐13 at 37°C for 6 hrs, and the cleavage fragments were separated on a 12% SDS ‐ PAGE gel and detected with a silver staining kit. The left lane shows the 140‐ and 100‐ kD a forms of the intact Ln‐5γ2 chain. MMP ‐2 generated an 80 kD Ln‐5γ2x fragment and a very faint 66 kD fragment. MMP ‐13 further generated low molecular weight fragments that were approximately 20 kD. ( B ) LC ‐ MS / MS detection showed that the 20 kD fragment generated by MMP ‐13 was part of the Ln‐5γ2 chain. ( C ) MMP ‐13 cleaved the Ln‐5γ2 chain at Lys‐Cys (540–541) and Arg‐Leu (694–695). This fragment belongs to the Ln‐5γ2 chain III and IV domain.

Techniques Used: Silver Staining, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, SDS Page, Generated, Molecular Weight

Cleavage of Ln‐5 by  MMP ‐2 or  MMP ‐13 affected  EGFR  and F‐actin expression. ( A ) The  WB  results show that treatment with  MMP ‐2 Ln‐5 cleavage fragments enhanced the  EGFR , F‐actin and  EGFR  downstream targets Raf,  ERK  and  AKT  protein levels in cells, but the levels were reduced in the  MMP ‐13‐Ln‐5 group. Vimentin and α‐tubulin levels were not obviously changed in either group. ( B ) Immunofluorescence staining was performed, and higher co‐expression of  EGFR  and F‐actin filaments was observed in the  MMP ‐2 + Ln‐5 group, and the arrangement of the F‐actin cytoskeleton was dramatically changed compared with that of the  MMP ‐13 + Ln‐5 group. The expression of vimentin and α‐tubulin was not different between these two treatment groups in both H460 and H661 cells.
Figure Legend Snippet: Cleavage of Ln‐5 by MMP ‐2 or MMP ‐13 affected EGFR and F‐actin expression. ( A ) The WB results show that treatment with MMP ‐2 Ln‐5 cleavage fragments enhanced the EGFR , F‐actin and EGFR downstream targets Raf, ERK and AKT protein levels in cells, but the levels were reduced in the MMP ‐13‐Ln‐5 group. Vimentin and α‐tubulin levels were not obviously changed in either group. ( B ) Immunofluorescence staining was performed, and higher co‐expression of EGFR and F‐actin filaments was observed in the MMP ‐2 + Ln‐5 group, and the arrangement of the F‐actin cytoskeleton was dramatically changed compared with that of the MMP ‐13 + Ln‐5 group. The expression of vimentin and α‐tubulin was not different between these two treatment groups in both H460 and H661 cells.

Techniques Used: Expressing, Western Blot, Immunofluorescence, Staining

VM  and the expression of  MMP ‐13,  MMP ‐2, Ln‐5 and  EGFR  in  LCLC  tissues. ( A ) Representative images of the presence of  VM  in human  LCLC  tissues ( CD 34/ PAS  double‐staining). The yellow arrow shows that the endothelial‐dependent vessel was positive for both  CD 34 and  PAS . The black arrows show  VM  channels in which  PAS ‐positive materials and red blood cells (red arrow) are observed. ( B )  VM ‐positive tissues exhibited significantly reduced  MMP ‐13 expression, and  VM ‐negative tissues exhibited significantly increased  MMP ‐13 expression ( P  = 0.005). In contrast,  VM ‐positive tissues exhibited significantly reduced  MMP ‐2 expression, and  VM ‐negative tissue exhibited significantly increased  MMP ‐2 expression. There was no significant correlation between  MMP ‐13 and  MMP ‐2 ( P  = 0.301). ( C ). Significantly reduced expression of Ln‐5 and  EGFR  can be observed in the  LCLC  samples with  MMP ‐13‐positive tissues compared with expression in  VM ‐negative tissues ( P
Figure Legend Snippet: VM and the expression of MMP ‐13, MMP ‐2, Ln‐5 and EGFR in LCLC tissues. ( A ) Representative images of the presence of VM in human LCLC tissues ( CD 34/ PAS double‐staining). The yellow arrow shows that the endothelial‐dependent vessel was positive for both CD 34 and PAS . The black arrows show VM channels in which PAS ‐positive materials and red blood cells (red arrow) are observed. ( B ) VM ‐positive tissues exhibited significantly reduced MMP ‐13 expression, and VM ‐negative tissues exhibited significantly increased MMP ‐13 expression ( P  = 0.005). In contrast, VM ‐positive tissues exhibited significantly reduced MMP ‐2 expression, and VM ‐negative tissue exhibited significantly increased MMP ‐2 expression. There was no significant correlation between MMP ‐13 and MMP ‐2 ( P  = 0.301). ( C ). Significantly reduced expression of Ln‐5 and EGFR can be observed in the LCLC samples with MMP ‐13‐positive tissues compared with expression in VM ‐negative tissues ( P

Techniques Used: Expressing, Double Staining

25) Product Images from "MMP‐2 and MMP‐13 affect vasculogenic mimicry formation in large cell lung cancer"

Article Title: MMP‐2 and MMP‐13 affect vasculogenic mimicry formation in large cell lung cancer

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.13283

MMP ‐13 and  MMP ‐2 expression in different tumour stages. ( A ) Tumour volume at different weeks. ( B ) Endomucin/ PAS  double‐staining in H460 xenografts showed decreased  VM  but increased  EDV  numbers (Bar = 100 μm). The black arrow shows  VM  formation. The red arrow shows the  EDV s. ( C ) The expression levels of  MMP ‐13 showed a gradually increasing trend with tumour growth. Meanwhile, no significant difference in the  MMP ‐2 expression was observed at different tumour growth stages (Bar = 100 μm).  WB  showed  MMP ‐13 gradually increasing, but Ln‐5 and  EGFR  showed a decreasing trend with tumour growth.  MMP ‐2 expression was observed at different tumour growth stages.
Figure Legend Snippet: MMP ‐13 and MMP ‐2 expression in different tumour stages. ( A ) Tumour volume at different weeks. ( B ) Endomucin/ PAS double‐staining in H460 xenografts showed decreased VM but increased EDV numbers (Bar = 100 μm). The black arrow shows VM formation. The red arrow shows the EDV s. ( C ) The expression levels of MMP ‐13 showed a gradually increasing trend with tumour growth. Meanwhile, no significant difference in the MMP ‐2 expression was observed at different tumour growth stages (Bar = 100 μm). WB showed MMP ‐13 gradually increasing, but Ln‐5 and EGFR showed a decreasing trend with tumour growth. MMP ‐2 expression was observed at different tumour growth stages.

Techniques Used: Expressing, Double Staining, Western Blot

Cleavage of Ln‐5 by  MMP ‐13 was detected with a silver staining kit and  LC ‐ MS / MS . ( A ) Human Ln‐5 was cleaved by  MMP ‐2 and  MMP ‐13 at 37°C for 6 hrs, and the cleavage fragments were separated on a 12%  SDS ‐ PAGE  gel and detected with a silver staining kit. The left lane shows the 140‐ and 100‐ kD a forms of the intact Ln‐5γ2 chain.  MMP ‐2 generated an 80 kD Ln‐5γ2x fragment and a very faint 66 kD fragment.  MMP ‐13 further generated low molecular weight fragments that were approximately 20 kD. ( B )  LC ‐ MS / MS  detection showed that the 20 kD fragment generated by  MMP ‐13 was part of the Ln‐5γ2 chain. ( C )  MMP ‐13 cleaved the Ln‐5γ2 chain at Lys‐Cys (540–541) and Arg‐Leu (694–695). This fragment belongs to the Ln‐5γ2 chain  III  and IV domain.
Figure Legend Snippet: Cleavage of Ln‐5 by MMP ‐13 was detected with a silver staining kit and LC ‐ MS / MS . ( A ) Human Ln‐5 was cleaved by MMP ‐2 and MMP ‐13 at 37°C for 6 hrs, and the cleavage fragments were separated on a 12% SDS ‐ PAGE gel and detected with a silver staining kit. The left lane shows the 140‐ and 100‐ kD a forms of the intact Ln‐5γ2 chain. MMP ‐2 generated an 80 kD Ln‐5γ2x fragment and a very faint 66 kD fragment. MMP ‐13 further generated low molecular weight fragments that were approximately 20 kD. ( B ) LC ‐ MS / MS detection showed that the 20 kD fragment generated by MMP ‐13 was part of the Ln‐5γ2 chain. ( C ) MMP ‐13 cleaved the Ln‐5γ2 chain at Lys‐Cys (540–541) and Arg‐Leu (694–695). This fragment belongs to the Ln‐5γ2 chain III and IV domain.

Techniques Used: Silver Staining, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, SDS Page, Generated, Molecular Weight

Cleavage of Ln‐5 by  MMP ‐2 or  MMP ‐13 affected  EGFR  and F‐actin expression. ( A ) The  WB  results show that treatment with  MMP ‐2 Ln‐5 cleavage fragments enhanced the  EGFR , F‐actin and  EGFR  downstream targets Raf,  ERK  and  AKT  protein levels in cells, but the levels were reduced in the  MMP ‐13‐Ln‐5 group. Vimentin and α‐tubulin levels were not obviously changed in either group. ( B ) Immunofluorescence staining was performed, and higher co‐expression of  EGFR  and F‐actin filaments was observed in the  MMP ‐2 + Ln‐5 group, and the arrangement of the F‐actin cytoskeleton was dramatically changed compared with that of the  MMP ‐13 + Ln‐5 group. The expression of vimentin and α‐tubulin was not different between these two treatment groups in both H460 and H661 cells.
Figure Legend Snippet: Cleavage of Ln‐5 by MMP ‐2 or MMP ‐13 affected EGFR and F‐actin expression. ( A ) The WB results show that treatment with MMP ‐2 Ln‐5 cleavage fragments enhanced the EGFR , F‐actin and EGFR downstream targets Raf, ERK and AKT protein levels in cells, but the levels were reduced in the MMP ‐13‐Ln‐5 group. Vimentin and α‐tubulin levels were not obviously changed in either group. ( B ) Immunofluorescence staining was performed, and higher co‐expression of EGFR and F‐actin filaments was observed in the MMP ‐2 + Ln‐5 group, and the arrangement of the F‐actin cytoskeleton was dramatically changed compared with that of the MMP ‐13 + Ln‐5 group. The expression of vimentin and α‐tubulin was not different between these two treatment groups in both H460 and H661 cells.

Techniques Used: Expressing, Western Blot, Immunofluorescence, Staining

VM  and the expression of  MMP ‐13,  MMP ‐2, Ln‐5 and  EGFR  in  LCLC  tissues. ( A ) Representative images of the presence of  VM  in human  LCLC  tissues ( CD 34/ PAS  double‐staining). The yellow arrow shows that the endothelial‐dependent vessel was positive for both  CD 34 and  PAS . The black arrows show  VM  channels in which  PAS ‐positive materials and red blood cells (red arrow) are observed. ( B )  VM ‐positive tissues exhibited significantly reduced  MMP ‐13 expression, and  VM ‐negative tissues exhibited significantly increased  MMP ‐13 expression ( P  = 0.005). In contrast,  VM ‐positive tissues exhibited significantly reduced  MMP ‐2 expression, and  VM ‐negative tissue exhibited significantly increased  MMP ‐2 expression. There was no significant correlation between  MMP ‐13 and  MMP ‐2 ( P  = 0.301). ( C ). Significantly reduced expression of Ln‐5 and  EGFR  can be observed in the  LCLC  samples with  MMP ‐13‐positive tissues compared with expression in  VM ‐negative tissues ( P
Figure Legend Snippet: VM and the expression of MMP ‐13, MMP ‐2, Ln‐5 and EGFR in LCLC tissues. ( A ) Representative images of the presence of VM in human LCLC tissues ( CD 34/ PAS double‐staining). The yellow arrow shows that the endothelial‐dependent vessel was positive for both CD 34 and PAS . The black arrows show VM channels in which PAS ‐positive materials and red blood cells (red arrow) are observed. ( B ) VM ‐positive tissues exhibited significantly reduced MMP ‐13 expression, and VM ‐negative tissues exhibited significantly increased MMP ‐13 expression ( P  = 0.005). In contrast, VM ‐positive tissues exhibited significantly reduced MMP ‐2 expression, and VM ‐negative tissue exhibited significantly increased MMP ‐2 expression. There was no significant correlation between MMP ‐13 and MMP ‐2 ( P  = 0.301). ( C ). Significantly reduced expression of Ln‐5 and EGFR can be observed in the LCLC samples with MMP ‐13‐positive tissues compared with expression in VM ‐negative tissues ( P

Techniques Used: Expressing, Double Staining

26) Product Images from "MMP‐2 and MMP‐13 affect vasculogenic mimicry formation in large cell lung cancer"

Article Title: MMP‐2 and MMP‐13 affect vasculogenic mimicry formation in large cell lung cancer

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.13283

MMP ‐13 and  MMP ‐2 expression in different tumour stages. ( A ) Tumour volume at different weeks. ( B ) Endomucin/ PAS  double‐staining in H460 xenografts showed decreased  VM  but increased  EDV  numbers (Bar = 100 μm). The black arrow shows  VM  formation. The red arrow shows the  EDV s. ( C ) The expression levels of  MMP ‐13 showed a gradually increasing trend with tumour growth. Meanwhile, no significant difference in the  MMP ‐2 expression was observed at different tumour growth stages (Bar = 100 μm).  WB  showed  MMP ‐13 gradually increasing, but Ln‐5 and  EGFR  showed a decreasing trend with tumour growth.  MMP ‐2 expression was observed at different tumour growth stages.
Figure Legend Snippet: MMP ‐13 and MMP ‐2 expression in different tumour stages. ( A ) Tumour volume at different weeks. ( B ) Endomucin/ PAS double‐staining in H460 xenografts showed decreased VM but increased EDV numbers (Bar = 100 μm). The black arrow shows VM formation. The red arrow shows the EDV s. ( C ) The expression levels of MMP ‐13 showed a gradually increasing trend with tumour growth. Meanwhile, no significant difference in the MMP ‐2 expression was observed at different tumour growth stages (Bar = 100 μm). WB showed MMP ‐13 gradually increasing, but Ln‐5 and EGFR showed a decreasing trend with tumour growth. MMP ‐2 expression was observed at different tumour growth stages.

Techniques Used: Expressing, Double Staining, Western Blot

Cleavage of Ln‐5 by  MMP ‐13 was detected with a silver staining kit and  LC ‐ MS / MS . ( A ) Human Ln‐5 was cleaved by  MMP ‐2 and  MMP ‐13 at 37°C for 6 hrs, and the cleavage fragments were separated on a 12%  SDS ‐ PAGE  gel and detected with a silver staining kit. The left lane shows the 140‐ and 100‐ kD a forms of the intact Ln‐5γ2 chain.  MMP ‐2 generated an 80 kD Ln‐5γ2x fragment and a very faint 66 kD fragment.  MMP ‐13 further generated low molecular weight fragments that were approximately 20 kD. ( B )  LC ‐ MS / MS  detection showed that the 20 kD fragment generated by  MMP ‐13 was part of the Ln‐5γ2 chain. ( C )  MMP ‐13 cleaved the Ln‐5γ2 chain at Lys‐Cys (540–541) and Arg‐Leu (694–695). This fragment belongs to the Ln‐5γ2 chain  III  and IV domain.
Figure Legend Snippet: Cleavage of Ln‐5 by MMP ‐13 was detected with a silver staining kit and LC ‐ MS / MS . ( A ) Human Ln‐5 was cleaved by MMP ‐2 and MMP ‐13 at 37°C for 6 hrs, and the cleavage fragments were separated on a 12% SDS ‐ PAGE gel and detected with a silver staining kit. The left lane shows the 140‐ and 100‐ kD a forms of the intact Ln‐5γ2 chain. MMP ‐2 generated an 80 kD Ln‐5γ2x fragment and a very faint 66 kD fragment. MMP ‐13 further generated low molecular weight fragments that were approximately 20 kD. ( B ) LC ‐ MS / MS detection showed that the 20 kD fragment generated by MMP ‐13 was part of the Ln‐5γ2 chain. ( C ) MMP ‐13 cleaved the Ln‐5γ2 chain at Lys‐Cys (540–541) and Arg‐Leu (694–695). This fragment belongs to the Ln‐5γ2 chain III and IV domain.

Techniques Used: Silver Staining, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, SDS Page, Generated, Molecular Weight

Cleavage of Ln‐5 by  MMP ‐2 or  MMP ‐13 affected  EGFR  and F‐actin expression. ( A ) The  WB  results show that treatment with  MMP ‐2 Ln‐5 cleavage fragments enhanced the  EGFR , F‐actin and  EGFR  downstream targets Raf,  ERK  and  AKT  protein levels in cells, but the levels were reduced in the  MMP ‐13‐Ln‐5 group. Vimentin and α‐tubulin levels were not obviously changed in either group. ( B ) Immunofluorescence staining was performed, and higher co‐expression of  EGFR  and F‐actin filaments was observed in the  MMP ‐2 + Ln‐5 group, and the arrangement of the F‐actin cytoskeleton was dramatically changed compared with that of the  MMP ‐13 + Ln‐5 group. The expression of vimentin and α‐tubulin was not different between these two treatment groups in both H460 and H661 cells.
Figure Legend Snippet: Cleavage of Ln‐5 by MMP ‐2 or MMP ‐13 affected EGFR and F‐actin expression. ( A ) The WB results show that treatment with MMP ‐2 Ln‐5 cleavage fragments enhanced the EGFR , F‐actin and EGFR downstream targets Raf, ERK and AKT protein levels in cells, but the levels were reduced in the MMP ‐13‐Ln‐5 group. Vimentin and α‐tubulin levels were not obviously changed in either group. ( B ) Immunofluorescence staining was performed, and higher co‐expression of EGFR and F‐actin filaments was observed in the MMP ‐2 + Ln‐5 group, and the arrangement of the F‐actin cytoskeleton was dramatically changed compared with that of the MMP ‐13 + Ln‐5 group. The expression of vimentin and α‐tubulin was not different between these two treatment groups in both H460 and H661 cells.

Techniques Used: Expressing, Western Blot, Immunofluorescence, Staining

VM  and the expression of  MMP ‐13,  MMP ‐2, Ln‐5 and  EGFR  in  LCLC  tissues. ( A ) Representative images of the presence of  VM  in human  LCLC  tissues ( CD 34/ PAS  double‐staining). The yellow arrow shows that the endothelial‐dependent vessel was positive for both  CD 34 and  PAS . The black arrows show  VM  channels in which  PAS ‐positive materials and red blood cells (red arrow) are observed. ( B )  VM ‐positive tissues exhibited significantly reduced  MMP ‐13 expression, and  VM ‐negative tissues exhibited significantly increased  MMP ‐13 expression ( P  = 0.005). In contrast,  VM ‐positive tissues exhibited significantly reduced  MMP ‐2 expression, and  VM ‐negative tissue exhibited significantly increased  MMP ‐2 expression. There was no significant correlation between  MMP ‐13 and  MMP ‐2 ( P  = 0.301). ( C ). Significantly reduced expression of Ln‐5 and  EGFR  can be observed in the  LCLC  samples with  MMP ‐13‐positive tissues compared with expression in  VM ‐negative tissues ( P
Figure Legend Snippet: VM and the expression of MMP ‐13, MMP ‐2, Ln‐5 and EGFR in LCLC tissues. ( A ) Representative images of the presence of VM in human LCLC tissues ( CD 34/ PAS double‐staining). The yellow arrow shows that the endothelial‐dependent vessel was positive for both CD 34 and PAS . The black arrows show VM channels in which PAS ‐positive materials and red blood cells (red arrow) are observed. ( B ) VM ‐positive tissues exhibited significantly reduced MMP ‐13 expression, and VM ‐negative tissues exhibited significantly increased MMP ‐13 expression ( P  = 0.005). In contrast, VM ‐positive tissues exhibited significantly reduced MMP ‐2 expression, and VM ‐negative tissue exhibited significantly increased MMP ‐2 expression. There was no significant correlation between MMP ‐13 and MMP ‐2 ( P  = 0.301). ( C ). Significantly reduced expression of Ln‐5 and EGFR can be observed in the LCLC samples with MMP ‐13‐positive tissues compared with expression in VM ‐negative tissues ( P

Techniques Used: Expressing, Double Staining

27) Product Images from "MMP‐2 and MMP‐13 affect vasculogenic mimicry formation in large cell lung cancer"

Article Title: MMP‐2 and MMP‐13 affect vasculogenic mimicry formation in large cell lung cancer

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.13283

MMP ‐13 and  MMP ‐2 expression in different tumour stages. ( A ) Tumour volume at different weeks. ( B ) Endomucin/ PAS  double‐staining in H460 xenografts showed decreased  VM  but increased  EDV  numbers (Bar = 100 μm). The black arrow shows  VM  formation. The red arrow shows the  EDV s. ( C ) The expression levels of  MMP ‐13 showed a gradually increasing trend with tumour growth. Meanwhile, no significant difference in the  MMP ‐2 expression was observed at different tumour growth stages (Bar = 100 μm).  WB  showed  MMP ‐13 gradually increasing, but Ln‐5 and  EGFR  showed a decreasing trend with tumour growth.  MMP ‐2 expression was observed at different tumour growth stages.
Figure Legend Snippet: MMP ‐13 and MMP ‐2 expression in different tumour stages. ( A ) Tumour volume at different weeks. ( B ) Endomucin/ PAS double‐staining in H460 xenografts showed decreased VM but increased EDV numbers (Bar = 100 μm). The black arrow shows VM formation. The red arrow shows the EDV s. ( C ) The expression levels of MMP ‐13 showed a gradually increasing trend with tumour growth. Meanwhile, no significant difference in the MMP ‐2 expression was observed at different tumour growth stages (Bar = 100 μm). WB showed MMP ‐13 gradually increasing, but Ln‐5 and EGFR showed a decreasing trend with tumour growth. MMP ‐2 expression was observed at different tumour growth stages.

Techniques Used: Expressing, Double Staining, Western Blot

Cleavage of Ln‐5 by  MMP ‐13 was detected with a silver staining kit and  LC ‐ MS / MS . ( A ) Human Ln‐5 was cleaved by  MMP ‐2 and  MMP ‐13 at 37°C for 6 hrs, and the cleavage fragments were separated on a 12%  SDS ‐ PAGE  gel and detected with a silver staining kit. The left lane shows the 140‐ and 100‐ kD a forms of the intact Ln‐5γ2 chain.  MMP ‐2 generated an 80 kD Ln‐5γ2x fragment and a very faint 66 kD fragment.  MMP ‐13 further generated low molecular weight fragments that were approximately 20 kD. ( B )  LC ‐ MS / MS  detection showed that the 20 kD fragment generated by  MMP ‐13 was part of the Ln‐5γ2 chain. ( C )  MMP ‐13 cleaved the Ln‐5γ2 chain at Lys‐Cys (540–541) and Arg‐Leu (694–695). This fragment belongs to the Ln‐5γ2 chain  III  and IV domain.
Figure Legend Snippet: Cleavage of Ln‐5 by MMP ‐13 was detected with a silver staining kit and LC ‐ MS / MS . ( A ) Human Ln‐5 was cleaved by MMP ‐2 and MMP ‐13 at 37°C for 6 hrs, and the cleavage fragments were separated on a 12% SDS ‐ PAGE gel and detected with a silver staining kit. The left lane shows the 140‐ and 100‐ kD a forms of the intact Ln‐5γ2 chain. MMP ‐2 generated an 80 kD Ln‐5γ2x fragment and a very faint 66 kD fragment. MMP ‐13 further generated low molecular weight fragments that were approximately 20 kD. ( B ) LC ‐ MS / MS detection showed that the 20 kD fragment generated by MMP ‐13 was part of the Ln‐5γ2 chain. ( C ) MMP ‐13 cleaved the Ln‐5γ2 chain at Lys‐Cys (540–541) and Arg‐Leu (694–695). This fragment belongs to the Ln‐5γ2 chain III and IV domain.

Techniques Used: Silver Staining, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, SDS Page, Generated, Molecular Weight

Cleavage of Ln‐5 by  MMP ‐2 or  MMP ‐13 affected  EGFR  and F‐actin expression. ( A ) The  WB  results show that treatment with  MMP ‐2 Ln‐5 cleavage fragments enhanced the  EGFR , F‐actin and  EGFR  downstream targets Raf,  ERK  and  AKT  protein levels in cells, but the levels were reduced in the  MMP ‐13‐Ln‐5 group. Vimentin and α‐tubulin levels were not obviously changed in either group. ( B ) Immunofluorescence staining was performed, and higher co‐expression of  EGFR  and F‐actin filaments was observed in the  MMP ‐2 + Ln‐5 group, and the arrangement of the F‐actin cytoskeleton was dramatically changed compared with that of the  MMP ‐13 + Ln‐5 group. The expression of vimentin and α‐tubulin was not different between these two treatment groups in both H460 and H661 cells.
Figure Legend Snippet: Cleavage of Ln‐5 by MMP ‐2 or MMP ‐13 affected EGFR and F‐actin expression. ( A ) The WB results show that treatment with MMP ‐2 Ln‐5 cleavage fragments enhanced the EGFR , F‐actin and EGFR downstream targets Raf, ERK and AKT protein levels in cells, but the levels were reduced in the MMP ‐13‐Ln‐5 group. Vimentin and α‐tubulin levels were not obviously changed in either group. ( B ) Immunofluorescence staining was performed, and higher co‐expression of EGFR and F‐actin filaments was observed in the MMP ‐2 + Ln‐5 group, and the arrangement of the F‐actin cytoskeleton was dramatically changed compared with that of the MMP ‐13 + Ln‐5 group. The expression of vimentin and α‐tubulin was not different between these two treatment groups in both H460 and H661 cells.

Techniques Used: Expressing, Western Blot, Immunofluorescence, Staining

VM  and the expression of  MMP ‐13,  MMP ‐2, Ln‐5 and  EGFR  in  LCLC  tissues. ( A ) Representative images of the presence of  VM  in human  LCLC  tissues ( CD 34/ PAS  double‐staining). The yellow arrow shows that the endothelial‐dependent vessel was positive for both  CD 34 and  PAS . The black arrows show  VM  channels in which  PAS ‐positive materials and red blood cells (red arrow) are observed. ( B )  VM ‐positive tissues exhibited significantly reduced  MMP ‐13 expression, and  VM ‐negative tissues exhibited significantly increased  MMP ‐13 expression ( P  = 0.005). In contrast,  VM ‐positive tissues exhibited significantly reduced  MMP ‐2 expression, and  VM ‐negative tissue exhibited significantly increased  MMP ‐2 expression. There was no significant correlation between  MMP ‐13 and  MMP ‐2 ( P  = 0.301). ( C ). Significantly reduced expression of Ln‐5 and  EGFR  can be observed in the  LCLC  samples with  MMP ‐13‐positive tissues compared with expression in  VM ‐negative tissues ( P
Figure Legend Snippet: VM and the expression of MMP ‐13, MMP ‐2, Ln‐5 and EGFR in LCLC tissues. ( A ) Representative images of the presence of VM in human LCLC tissues ( CD 34/ PAS double‐staining). The yellow arrow shows that the endothelial‐dependent vessel was positive for both CD 34 and PAS . The black arrows show VM channels in which PAS ‐positive materials and red blood cells (red arrow) are observed. ( B ) VM ‐positive tissues exhibited significantly reduced MMP ‐13 expression, and VM ‐negative tissues exhibited significantly increased MMP ‐13 expression ( P  = 0.005). In contrast, VM ‐positive tissues exhibited significantly reduced MMP ‐2 expression, and VM ‐negative tissue exhibited significantly increased MMP ‐2 expression. There was no significant correlation between MMP ‐13 and MMP ‐2 ( P  = 0.301). ( C ). Significantly reduced expression of Ln‐5 and EGFR can be observed in the LCLC samples with MMP ‐13‐positive tissues compared with expression in VM ‐negative tissues ( P

Techniques Used: Expressing, Double Staining

28) Product Images from "MMP‐2 and MMP‐13 affect vasculogenic mimicry formation in large cell lung cancer"

Article Title: MMP‐2 and MMP‐13 affect vasculogenic mimicry formation in large cell lung cancer

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.13283

MMP ‐13 and  MMP ‐2 expression in different tumour stages. ( A ) Tumour volume at different weeks. ( B ) Endomucin/ PAS  double‐staining in H460 xenografts showed decreased  VM  but increased  EDV  numbers (Bar = 100 μm). The black arrow shows  VM  formation. The red arrow shows the  EDV s. ( C ) The expression levels of  MMP ‐13 showed a gradually increasing trend with tumour growth. Meanwhile, no significant difference in the  MMP ‐2 expression was observed at different tumour growth stages (Bar = 100 μm).  WB  showed  MMP ‐13 gradually increasing, but Ln‐5 and  EGFR  showed a decreasing trend with tumour growth.  MMP ‐2 expression was observed at different tumour growth stages.
Figure Legend Snippet: MMP ‐13 and MMP ‐2 expression in different tumour stages. ( A ) Tumour volume at different weeks. ( B ) Endomucin/ PAS double‐staining in H460 xenografts showed decreased VM but increased EDV numbers (Bar = 100 μm). The black arrow shows VM formation. The red arrow shows the EDV s. ( C ) The expression levels of MMP ‐13 showed a gradually increasing trend with tumour growth. Meanwhile, no significant difference in the MMP ‐2 expression was observed at different tumour growth stages (Bar = 100 μm). WB showed MMP ‐13 gradually increasing, but Ln‐5 and EGFR showed a decreasing trend with tumour growth. MMP ‐2 expression was observed at different tumour growth stages.

Techniques Used: Expressing, Double Staining, Western Blot

Cleavage of Ln‐5 by  MMP ‐13 was detected with a silver staining kit and  LC ‐ MS / MS . ( A ) Human Ln‐5 was cleaved by  MMP ‐2 and  MMP ‐13 at 37°C for 6 hrs, and the cleavage fragments were separated on a 12%  SDS ‐ PAGE  gel and detected with a silver staining kit. The left lane shows the 140‐ and 100‐ kD a forms of the intact Ln‐5γ2 chain.  MMP ‐2 generated an 80 kD Ln‐5γ2x fragment and a very faint 66 kD fragment.  MMP ‐13 further generated low molecular weight fragments that were approximately 20 kD. ( B )  LC ‐ MS / MS  detection showed that the 20 kD fragment generated by  MMP ‐13 was part of the Ln‐5γ2 chain. ( C )  MMP ‐13 cleaved the Ln‐5γ2 chain at Lys‐Cys (540–541) and Arg‐Leu (694–695). This fragment belongs to the Ln‐5γ2 chain  III  and IV domain.
Figure Legend Snippet: Cleavage of Ln‐5 by MMP ‐13 was detected with a silver staining kit and LC ‐ MS / MS . ( A ) Human Ln‐5 was cleaved by MMP ‐2 and MMP ‐13 at 37°C for 6 hrs, and the cleavage fragments were separated on a 12% SDS ‐ PAGE gel and detected with a silver staining kit. The left lane shows the 140‐ and 100‐ kD a forms of the intact Ln‐5γ2 chain. MMP ‐2 generated an 80 kD Ln‐5γ2x fragment and a very faint 66 kD fragment. MMP ‐13 further generated low molecular weight fragments that were approximately 20 kD. ( B ) LC ‐ MS / MS detection showed that the 20 kD fragment generated by MMP ‐13 was part of the Ln‐5γ2 chain. ( C ) MMP ‐13 cleaved the Ln‐5γ2 chain at Lys‐Cys (540–541) and Arg‐Leu (694–695). This fragment belongs to the Ln‐5γ2 chain III and IV domain.

Techniques Used: Silver Staining, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, SDS Page, Generated, Molecular Weight

Cleavage of Ln‐5 by  MMP ‐2 or  MMP ‐13 affected  EGFR  and F‐actin expression. ( A ) The  WB  results show that treatment with  MMP ‐2 Ln‐5 cleavage fragments enhanced the  EGFR , F‐actin and  EGFR  downstream targets Raf,  ERK  and  AKT  protein levels in cells, but the levels were reduced in the  MMP ‐13‐Ln‐5 group. Vimentin and α‐tubulin levels were not obviously changed in either group. ( B ) Immunofluorescence staining was performed, and higher co‐expression of  EGFR  and F‐actin filaments was observed in the  MMP ‐2 + Ln‐5 group, and the arrangement of the F‐actin cytoskeleton was dramatically changed compared with that of the  MMP ‐13 + Ln‐5 group. The expression of vimentin and α‐tubulin was not different between these two treatment groups in both H460 and H661 cells.
Figure Legend Snippet: Cleavage of Ln‐5 by MMP ‐2 or MMP ‐13 affected EGFR and F‐actin expression. ( A ) The WB results show that treatment with MMP ‐2 Ln‐5 cleavage fragments enhanced the EGFR , F‐actin and EGFR downstream targets Raf, ERK and AKT protein levels in cells, but the levels were reduced in the MMP ‐13‐Ln‐5 group. Vimentin and α‐tubulin levels were not obviously changed in either group. ( B ) Immunofluorescence staining was performed, and higher co‐expression of EGFR and F‐actin filaments was observed in the MMP ‐2 + Ln‐5 group, and the arrangement of the F‐actin cytoskeleton was dramatically changed compared with that of the MMP ‐13 + Ln‐5 group. The expression of vimentin and α‐tubulin was not different between these two treatment groups in both H460 and H661 cells.

Techniques Used: Expressing, Western Blot, Immunofluorescence, Staining

VM  and the expression of  MMP ‐13,  MMP ‐2, Ln‐5 and  EGFR  in  LCLC  tissues. ( A ) Representative images of the presence of  VM  in human  LCLC  tissues ( CD 34/ PAS  double‐staining). The yellow arrow shows that the endothelial‐dependent vessel was positive for both  CD 34 and  PAS . The black arrows show  VM  channels in which  PAS ‐positive materials and red blood cells (red arrow) are observed. ( B )  VM ‐positive tissues exhibited significantly reduced  MMP ‐13 expression, and  VM ‐negative tissues exhibited significantly increased  MMP ‐13 expression ( P  = 0.005). In contrast,  VM ‐positive tissues exhibited significantly reduced  MMP ‐2 expression, and  VM ‐negative tissue exhibited significantly increased  MMP ‐2 expression. There was no significant correlation between  MMP ‐13 and  MMP ‐2 ( P  = 0.301). ( C ). Significantly reduced expression of Ln‐5 and  EGFR  can be observed in the  LCLC  samples with  MMP ‐13‐positive tissues compared with expression in  VM ‐negative tissues ( P
Figure Legend Snippet: VM and the expression of MMP ‐13, MMP ‐2, Ln‐5 and EGFR in LCLC tissues. ( A ) Representative images of the presence of VM in human LCLC tissues ( CD 34/ PAS double‐staining). The yellow arrow shows that the endothelial‐dependent vessel was positive for both CD 34 and PAS . The black arrows show VM channels in which PAS ‐positive materials and red blood cells (red arrow) are observed. ( B ) VM ‐positive tissues exhibited significantly reduced MMP ‐13 expression, and VM ‐negative tissues exhibited significantly increased MMP ‐13 expression ( P  = 0.005). In contrast, VM ‐positive tissues exhibited significantly reduced MMP ‐2 expression, and VM ‐negative tissue exhibited significantly increased MMP ‐2 expression. There was no significant correlation between MMP ‐13 and MMP ‐2 ( P  = 0.301). ( C ). Significantly reduced expression of Ln‐5 and EGFR can be observed in the LCLC samples with MMP ‐13‐positive tissues compared with expression in VM ‐negative tissues ( P

Techniques Used: Expressing, Double Staining

29) Product Images from "MMP‐2 and MMP‐13 affect vasculogenic mimicry formation in large cell lung cancer"

Article Title: MMP‐2 and MMP‐13 affect vasculogenic mimicry formation in large cell lung cancer

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.13283

MMP ‐13 and  MMP ‐2 expression in different tumour stages. ( A ) Tumour volume at different weeks. ( B ) Endomucin/ PAS  double‐staining in H460 xenografts showed decreased  VM  but increased  EDV  numbers (Bar = 100 μm). The black arrow shows  VM  formation. The red arrow shows the  EDV s. ( C ) The expression levels of  MMP ‐13 showed a gradually increasing trend with tumour growth. Meanwhile, no significant difference in the  MMP ‐2 expression was observed at different tumour growth stages (Bar = 100 μm).  WB  showed  MMP ‐13 gradually increasing, but Ln‐5 and  EGFR  showed a decreasing trend with tumour growth.  MMP ‐2 expression was observed at different tumour growth stages.
Figure Legend Snippet: MMP ‐13 and MMP ‐2 expression in different tumour stages. ( A ) Tumour volume at different weeks. ( B ) Endomucin/ PAS double‐staining in H460 xenografts showed decreased VM but increased EDV numbers (Bar = 100 μm). The black arrow shows VM formation. The red arrow shows the EDV s. ( C ) The expression levels of MMP ‐13 showed a gradually increasing trend with tumour growth. Meanwhile, no significant difference in the MMP ‐2 expression was observed at different tumour growth stages (Bar = 100 μm). WB showed MMP ‐13 gradually increasing, but Ln‐5 and EGFR showed a decreasing trend with tumour growth. MMP ‐2 expression was observed at different tumour growth stages.

Techniques Used: Expressing, Double Staining, Western Blot

Cleavage of Ln‐5 by  MMP ‐13 was detected with a silver staining kit and  LC ‐ MS / MS . ( A ) Human Ln‐5 was cleaved by  MMP ‐2 and  MMP ‐13 at 37°C for 6 hrs, and the cleavage fragments were separated on a 12%  SDS ‐ PAGE  gel and detected with a silver staining kit. The left lane shows the 140‐ and 100‐ kD a forms of the intact Ln‐5γ2 chain.  MMP ‐2 generated an 80 kD Ln‐5γ2x fragment and a very faint 66 kD fragment.  MMP ‐13 further generated low molecular weight fragments that were approximately 20 kD. ( B )  LC ‐ MS / MS  detection showed that the 20 kD fragment generated by  MMP ‐13 was part of the Ln‐5γ2 chain. ( C )  MMP ‐13 cleaved the Ln‐5γ2 chain at Lys‐Cys (540–541) and Arg‐Leu (694–695). This fragment belongs to the Ln‐5γ2 chain  III  and IV domain.
Figure Legend Snippet: Cleavage of Ln‐5 by MMP ‐13 was detected with a silver staining kit and LC ‐ MS / MS . ( A ) Human Ln‐5 was cleaved by MMP ‐2 and MMP ‐13 at 37°C for 6 hrs, and the cleavage fragments were separated on a 12% SDS ‐ PAGE gel and detected with a silver staining kit. The left lane shows the 140‐ and 100‐ kD a forms of the intact Ln‐5γ2 chain. MMP ‐2 generated an 80 kD Ln‐5γ2x fragment and a very faint 66 kD fragment. MMP ‐13 further generated low molecular weight fragments that were approximately 20 kD. ( B ) LC ‐ MS / MS detection showed that the 20 kD fragment generated by MMP ‐13 was part of the Ln‐5γ2 chain. ( C ) MMP ‐13 cleaved the Ln‐5γ2 chain at Lys‐Cys (540–541) and Arg‐Leu (694–695). This fragment belongs to the Ln‐5γ2 chain III and IV domain.

Techniques Used: Silver Staining, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, SDS Page, Generated, Molecular Weight

Cleavage of Ln‐5 by  MMP ‐2 or  MMP ‐13 affected  EGFR  and F‐actin expression. ( A ) The  WB  results show that treatment with  MMP ‐2 Ln‐5 cleavage fragments enhanced the  EGFR , F‐actin and  EGFR  downstream targets Raf,  ERK  and  AKT  protein levels in cells, but the levels were reduced in the  MMP ‐13‐Ln‐5 group. Vimentin and α‐tubulin levels were not obviously changed in either group. ( B ) Immunofluorescence staining was performed, and higher co‐expression of  EGFR  and F‐actin filaments was observed in the  MMP ‐2 + Ln‐5 group, and the arrangement of the F‐actin cytoskeleton was dramatically changed compared with that of the  MMP ‐13 + Ln‐5 group. The expression of vimentin and α‐tubulin was not different between these two treatment groups in both H460 and H661 cells.
Figure Legend Snippet: Cleavage of Ln‐5 by MMP ‐2 or MMP ‐13 affected EGFR and F‐actin expression. ( A ) The WB results show that treatment with MMP ‐2 Ln‐5 cleavage fragments enhanced the EGFR , F‐actin and EGFR downstream targets Raf, ERK and AKT protein levels in cells, but the levels were reduced in the MMP ‐13‐Ln‐5 group. Vimentin and α‐tubulin levels were not obviously changed in either group. ( B ) Immunofluorescence staining was performed, and higher co‐expression of EGFR and F‐actin filaments was observed in the MMP ‐2 + Ln‐5 group, and the arrangement of the F‐actin cytoskeleton was dramatically changed compared with that of the MMP ‐13 + Ln‐5 group. The expression of vimentin and α‐tubulin was not different between these two treatment groups in both H460 and H661 cells.

Techniques Used: Expressing, Western Blot, Immunofluorescence, Staining

VM  and the expression of  MMP ‐13,  MMP ‐2, Ln‐5 and  EGFR  in  LCLC  tissues. ( A ) Representative images of the presence of  VM  in human  LCLC  tissues ( CD 34/ PAS  double‐staining). The yellow arrow shows that the endothelial‐dependent vessel was positive for both  CD 34 and  PAS . The black arrows show  VM  channels in which  PAS ‐positive materials and red blood cells (red arrow) are observed. ( B )  VM ‐positive tissues exhibited significantly reduced  MMP ‐13 expression, and  VM ‐negative tissues exhibited significantly increased  MMP ‐13 expression ( P  = 0.005). In contrast,  VM ‐positive tissues exhibited significantly reduced  MMP ‐2 expression, and  VM ‐negative tissue exhibited significantly increased  MMP ‐2 expression. There was no significant correlation between  MMP ‐13 and  MMP ‐2 ( P  = 0.301). ( C ). Significantly reduced expression of Ln‐5 and  EGFR  can be observed in the  LCLC  samples with  MMP ‐13‐positive tissues compared with expression in  VM ‐negative tissues ( P
Figure Legend Snippet: VM and the expression of MMP ‐13, MMP ‐2, Ln‐5 and EGFR in LCLC tissues. ( A ) Representative images of the presence of VM in human LCLC tissues ( CD 34/ PAS double‐staining). The yellow arrow shows that the endothelial‐dependent vessel was positive for both CD 34 and PAS . The black arrows show VM channels in which PAS ‐positive materials and red blood cells (red arrow) are observed. ( B ) VM ‐positive tissues exhibited significantly reduced MMP ‐13 expression, and VM ‐negative tissues exhibited significantly increased MMP ‐13 expression ( P  = 0.005). In contrast, VM ‐positive tissues exhibited significantly reduced MMP ‐2 expression, and VM ‐negative tissue exhibited significantly increased MMP ‐2 expression. There was no significant correlation between MMP ‐13 and MMP ‐2 ( P  = 0.301). ( C ). Significantly reduced expression of Ln‐5 and EGFR can be observed in the LCLC samples with MMP ‐13‐positive tissues compared with expression in VM ‐negative tissues ( P

Techniques Used: Expressing, Double Staining

30) Product Images from "MMP‐2 and MMP‐13 affect vasculogenic mimicry formation in large cell lung cancer"

Article Title: MMP‐2 and MMP‐13 affect vasculogenic mimicry formation in large cell lung cancer

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/jcmm.13283

MMP ‐13 and  MMP ‐2 expression in different tumour stages. ( A ) Tumour volume at different weeks. ( B ) Endomucin/ PAS  double‐staining in H460 xenografts showed decreased  VM  but increased  EDV  numbers (Bar = 100 μm). The black arrow shows  VM  formation. The red arrow shows the  EDV s. ( C ) The expression levels of  MMP ‐13 showed a gradually increasing trend with tumour growth. Meanwhile, no significant difference in the  MMP ‐2 expression was observed at different tumour growth stages (Bar = 100 μm).  WB  showed  MMP ‐13 gradually increasing, but Ln‐5 and  EGFR  showed a decreasing trend with tumour growth.  MMP ‐2 expression was observed at different tumour growth stages.
Figure Legend Snippet: MMP ‐13 and MMP ‐2 expression in different tumour stages. ( A ) Tumour volume at different weeks. ( B ) Endomucin/ PAS double‐staining in H460 xenografts showed decreased VM but increased EDV numbers (Bar = 100 μm). The black arrow shows VM formation. The red arrow shows the EDV s. ( C ) The expression levels of MMP ‐13 showed a gradually increasing trend with tumour growth. Meanwhile, no significant difference in the MMP ‐2 expression was observed at different tumour growth stages (Bar = 100 μm). WB showed MMP ‐13 gradually increasing, but Ln‐5 and EGFR showed a decreasing trend with tumour growth. MMP ‐2 expression was observed at different tumour growth stages.

Techniques Used: Expressing, Double Staining, Western Blot

Cleavage of Ln‐5 by  MMP ‐13 was detected with a silver staining kit and  LC ‐ MS / MS . ( A ) Human Ln‐5 was cleaved by  MMP ‐2 and  MMP ‐13 at 37°C for 6 hrs, and the cleavage fragments were separated on a 12%  SDS ‐ PAGE  gel and detected with a silver staining kit. The left lane shows the 140‐ and 100‐ kD a forms of the intact Ln‐5γ2 chain.  MMP ‐2 generated an 80 kD Ln‐5γ2x fragment and a very faint 66 kD fragment.  MMP ‐13 further generated low molecular weight fragments that were approximately 20 kD. ( B )  LC ‐ MS / MS  detection showed that the 20 kD fragment generated by  MMP ‐13 was part of the Ln‐5γ2 chain. ( C )  MMP ‐13 cleaved the Ln‐5γ2 chain at Lys‐Cys (540–541) and Arg‐Leu (694–695). This fragment belongs to the Ln‐5γ2 chain  III  and IV domain.
Figure Legend Snippet: Cleavage of Ln‐5 by MMP ‐13 was detected with a silver staining kit and LC ‐ MS / MS . ( A ) Human Ln‐5 was cleaved by MMP ‐2 and MMP ‐13 at 37°C for 6 hrs, and the cleavage fragments were separated on a 12% SDS ‐ PAGE gel and detected with a silver staining kit. The left lane shows the 140‐ and 100‐ kD a forms of the intact Ln‐5γ2 chain. MMP ‐2 generated an 80 kD Ln‐5γ2x fragment and a very faint 66 kD fragment. MMP ‐13 further generated low molecular weight fragments that were approximately 20 kD. ( B ) LC ‐ MS / MS detection showed that the 20 kD fragment generated by MMP ‐13 was part of the Ln‐5γ2 chain. ( C ) MMP ‐13 cleaved the Ln‐5γ2 chain at Lys‐Cys (540–541) and Arg‐Leu (694–695). This fragment belongs to the Ln‐5γ2 chain III and IV domain.

Techniques Used: Silver Staining, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, SDS Page, Generated, Molecular Weight

Cleavage of Ln‐5 by  MMP ‐2 or  MMP ‐13 affected  EGFR  and F‐actin expression. ( A ) The  WB  results show that treatment with  MMP ‐2 Ln‐5 cleavage fragments enhanced the  EGFR , F‐actin and  EGFR  downstream targets Raf,  ERK  and  AKT  protein levels in cells, but the levels were reduced in the  MMP ‐13‐Ln‐5 group. Vimentin and α‐tubulin levels were not obviously changed in either group. ( B ) Immunofluorescence staining was performed, and higher co‐expression of  EGFR  and F‐actin filaments was observed in the  MMP ‐2 + Ln‐5 group, and the arrangement of the F‐actin cytoskeleton was dramatically changed compared with that of the  MMP ‐13 + Ln‐5 group. The expression of vimentin and α‐tubulin was not different between these two treatment groups in both H460 and H661 cells.
Figure Legend Snippet: Cleavage of Ln‐5 by MMP ‐2 or MMP ‐13 affected EGFR and F‐actin expression. ( A ) The WB results show that treatment with MMP ‐2 Ln‐5 cleavage fragments enhanced the EGFR , F‐actin and EGFR downstream targets Raf, ERK and AKT protein levels in cells, but the levels were reduced in the MMP ‐13‐Ln‐5 group. Vimentin and α‐tubulin levels were not obviously changed in either group. ( B ) Immunofluorescence staining was performed, and higher co‐expression of EGFR and F‐actin filaments was observed in the MMP ‐2 + Ln‐5 group, and the arrangement of the F‐actin cytoskeleton was dramatically changed compared with that of the MMP ‐13 + Ln‐5 group. The expression of vimentin and α‐tubulin was not different between these two treatment groups in both H460 and H661 cells.

Techniques Used: Expressing, Western Blot, Immunofluorescence, Staining

VM  and the expression of  MMP ‐13,  MMP ‐2, Ln‐5 and  EGFR  in  LCLC  tissues. ( A ) Representative images of the presence of  VM  in human  LCLC  tissues ( CD 34/ PAS  double‐staining). The yellow arrow shows that the endothelial‐dependent vessel was positive for both  CD 34 and  PAS . The black arrows show  VM  channels in which  PAS ‐positive materials and red blood cells (red arrow) are observed. ( B )  VM ‐positive tissues exhibited significantly reduced  MMP ‐13 expression, and  VM ‐negative tissues exhibited significantly increased  MMP ‐13 expression ( P  = 0.005). In contrast,  VM ‐positive tissues exhibited significantly reduced  MMP ‐2 expression, and  VM ‐negative tissue exhibited significantly increased  MMP ‐2 expression. There was no significant correlation between  MMP ‐13 and  MMP ‐2 ( P  = 0.301). ( C ). Significantly reduced expression of Ln‐5 and  EGFR  can be observed in the  LCLC  samples with  MMP ‐13‐positive tissues compared with expression in  VM ‐negative tissues ( P
Figure Legend Snippet: VM and the expression of MMP ‐13, MMP ‐2, Ln‐5 and EGFR in LCLC tissues. ( A ) Representative images of the presence of VM in human LCLC tissues ( CD 34/ PAS double‐staining). The yellow arrow shows that the endothelial‐dependent vessel was positive for both CD 34 and PAS . The black arrows show VM channels in which PAS ‐positive materials and red blood cells (red arrow) are observed. ( B ) VM ‐positive tissues exhibited significantly reduced MMP ‐13 expression, and VM ‐negative tissues exhibited significantly increased MMP ‐13 expression ( P  = 0.005). In contrast, VM ‐positive tissues exhibited significantly reduced MMP ‐2 expression, and VM ‐negative tissue exhibited significantly increased MMP ‐2 expression. There was no significant correlation between MMP ‐13 and MMP ‐2 ( P  = 0.301). ( C ). Significantly reduced expression of Ln‐5 and EGFR can be observed in the LCLC samples with MMP ‐13‐positive tissues compared with expression in VM ‐negative tissues ( P

Techniques Used: Expressing, Double Staining

31) Product Images from "TGF-β inhibits alveolar protein transport by promoting shedding, regulated intramembrane proteolysis, and transcriptional downregulation of megalin"

Article Title: TGF-β inhibits alveolar protein transport by promoting shedding, regulated intramembrane proteolysis, and transcriptional downregulation of megalin

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

doi: 10.1152/ajplung.00569.2016

Megalin interacts with MMP-2 and MMP-14 at the plasma membrane. RLE-6TN cells were treated with TGF-β (20 ng/ml) for 10 h and crude plasma membrane fractions were obtained by centrifugation from whole cell lysates. Proteins from the crude plasma membrane fraction were extracted and immunoprecipitation (IP) with specific antibodies for megalin performed. Co-IP was assessed by immunoblotting with specific antibodies for MMP-2 ( A ) or MMP-14 ( B ). Reverse co-IP experiments were also performed as described above but with an IP with MMP-2 ( C ) or MMP-14 ( D ) specific antibodies. Co-IP was assessed by immunoblotting with a specific antibody for megalin. Representative blots are shown; n = 5.
Figure Legend Snippet: Megalin interacts with MMP-2 and MMP-14 at the plasma membrane. RLE-6TN cells were treated with TGF-β (20 ng/ml) for 10 h and crude plasma membrane fractions were obtained by centrifugation from whole cell lysates. Proteins from the crude plasma membrane fraction were extracted and immunoprecipitation (IP) with specific antibodies for megalin performed. Co-IP was assessed by immunoblotting with specific antibodies for MMP-2 ( A ) or MMP-14 ( B ). Reverse co-IP experiments were also performed as described above but with an IP with MMP-2 ( C ) or MMP-14 ( D ) specific antibodies. Co-IP was assessed by immunoblotting with a specific antibody for megalin. Representative blots are shown; n = 5.

Techniques Used: Centrifugation, Immunoprecipitation, Co-Immunoprecipitation Assay

Silencing of MMP-2, -9, or -14 prevents TGF-β-induced reduction of megalin cell surface abundance and restores albumin binding and uptake. RLE-6TN cells were transfected with MMP-2 ( A ), -9 ( B ), or -14 ( C ) siRNA for 72 h and treated with TGF-β (20 ng/ml) for 10 h. Megalin cell surface stability was measured by biotin-streptavidin pulldown assay and detected by SDS-PAGE and IB. D : RLE-6TN cells were treated as described above and subsequently incubated with FITC-albumin, cell surface-bound and taken-up fractions were collected, and FITC fluorescence was quantified. Fluorescence readouts were normalized to total protein amount from the taken-up fraction. Results are shown as means ± SE. Two-way ANOVA and Šidák’s multiple comparisons, * P
Figure Legend Snippet: Silencing of MMP-2, -9, or -14 prevents TGF-β-induced reduction of megalin cell surface abundance and restores albumin binding and uptake. RLE-6TN cells were transfected with MMP-2 ( A ), -9 ( B ), or -14 ( C ) siRNA for 72 h and treated with TGF-β (20 ng/ml) for 10 h. Megalin cell surface stability was measured by biotin-streptavidin pulldown assay and detected by SDS-PAGE and IB. D : RLE-6TN cells were treated as described above and subsequently incubated with FITC-albumin, cell surface-bound and taken-up fractions were collected, and FITC fluorescence was quantified. Fluorescence readouts were normalized to total protein amount from the taken-up fraction. Results are shown as means ± SE. Two-way ANOVA and Šidák’s multiple comparisons, * P

Techniques Used: Binding Assay, Transfection, SDS Page, Incubation, Fluorescence

TGF-β regulates intracellular expression of MMP-2, -9, and -14. RLE-6TN cells were treated with TGF-β (20 ng/ml) up to 48 h. Whole cell homogenates were processed by SDS-PAGE and IB for MMP-2 ( A ), MMP-9 ( B ), and MMP-14 ( C ). Representative blots are shown. Results are shown as means ± SE. One-way ANOVA and Šidák’s or Dunnett’s multiple comparisons, * P
Figure Legend Snippet: TGF-β regulates intracellular expression of MMP-2, -9, and -14. RLE-6TN cells were treated with TGF-β (20 ng/ml) up to 48 h. Whole cell homogenates were processed by SDS-PAGE and IB for MMP-2 ( A ), MMP-9 ( B ), and MMP-14 ( C ). Representative blots are shown. Results are shown as means ± SE. One-way ANOVA and Šidák’s or Dunnett’s multiple comparisons, * P

Techniques Used: Expressing, SDS Page

TGF-β regulates extracellular activation of MMP-2 and promotes shedding of megalin ectodomain. RLE-6TN cells were treated with TGF-β (20 ng/ml) up to 48 h. Supernatants (SN) were collected, centrifuged to remove debris, and concentrated for subsequent analysis. A : specific ELISA for MMP-2 or MMP-9. Chemiluminescent units were normalized to protein concentration in the SN. Boxes represent the median + quartiles; the ends of the whiskers show the minimum and maximum of all data. Two-way ANOVA and Šidák’s multiple comparisons, ** P
Figure Legend Snippet: TGF-β regulates extracellular activation of MMP-2 and promotes shedding of megalin ectodomain. RLE-6TN cells were treated with TGF-β (20 ng/ml) up to 48 h. Supernatants (SN) were collected, centrifuged to remove debris, and concentrated for subsequent analysis. A : specific ELISA for MMP-2 or MMP-9. Chemiluminescent units were normalized to protein concentration in the SN. Boxes represent the median + quartiles; the ends of the whiskers show the minimum and maximum of all data. Two-way ANOVA and Šidák’s multiple comparisons, ** P

Techniques Used: Activation Assay, Enzyme-linked Immunosorbent Assay, Protein Concentration

32) Product Images from "CXCR3 activation by lentivirus infection suppresses neuronal autophagy: neuroprotective effects of antiretroviral therapy"

Article Title: CXCR3 activation by lentivirus infection suppresses neuronal autophagy: neuroprotective effects of antiretroviral therapy

Journal: The FASEB Journal

doi: 10.1096/fj.08-128819

Brain expression of CXCL12, MMP-2, and CXCR3 in FIV infection. A ) Immunoblotting showed increased full-length (1-67) and cleaved (5-67) CXCL12 in brains of FIV − compared with mock-infected animals. B ) CXCL12, CXCL12(5-67), MMP-2, and CXCR3 immunoreactivities were increased in brains of FIV + animals but were suppressed by concurrent ddI treatment. C ) MMP-2 immunoreactivity (IR) showed minimal induction in FIV + animals with and without ddI treatment. D , E ) CXCL12(1-67) ( D ) and CXCL12(5-67) ( E ) were induced by FIV infection, but these changes were reversed by ddI treatment. F ) CXCR3 showed minimal induction of both immunoreactive bands in FIV infection and was not affected by ddI treatment. * P
Figure Legend Snippet: Brain expression of CXCL12, MMP-2, and CXCR3 in FIV infection. A ) Immunoblotting showed increased full-length (1-67) and cleaved (5-67) CXCL12 in brains of FIV − compared with mock-infected animals. B ) CXCL12, CXCL12(5-67), MMP-2, and CXCR3 immunoreactivities were increased in brains of FIV + animals but were suppressed by concurrent ddI treatment. C ) MMP-2 immunoreactivity (IR) showed minimal induction in FIV + animals with and without ddI treatment. D , E ) CXCL12(1-67) ( D ) and CXCL12(5-67) ( E ) were induced by FIV infection, but these changes were reversed by ddI treatment. F ) CXCR3 showed minimal induction of both immunoreactive bands in FIV infection and was not affected by ddI treatment. * P

Techniques Used: Expressing, Infection

Neurobehavioral performance, viral burden, and neuroinflammation in FIV infection. A ) FIV-infected animals (FIV+) exhibit greater neurobehavioral deficits than mock-infected controls (FIV−), while ddI treatment reversed neurobehavioral deficits in FIV-infected animals (FIV+/ddI) and had no effects on mock-infected animals. B ) ddI treatment suppressed viral load in plasma of FIV-infected animals. C ) Glycoprotein, F4/80 , expressed in activated myeloid cells showed increased transcript abundance in FIV-infected animals, which was reversed by ddI treatment. D ) CXCL12 transcript levels were higher in FIV-infected animals but were not reduced by ddI treatment. E ) MMP-2 transcripts were unregulated in FIV-infected animals but were suppressed with ddI treatment. F ) GFAP transcriptional activity was not affected by FIV infection or ddI treatment. * P
Figure Legend Snippet: Neurobehavioral performance, viral burden, and neuroinflammation in FIV infection. A ) FIV-infected animals (FIV+) exhibit greater neurobehavioral deficits than mock-infected controls (FIV−), while ddI treatment reversed neurobehavioral deficits in FIV-infected animals (FIV+/ddI) and had no effects on mock-infected animals. B ) ddI treatment suppressed viral load in plasma of FIV-infected animals. C ) Glycoprotein, F4/80 , expressed in activated myeloid cells showed increased transcript abundance in FIV-infected animals, which was reversed by ddI treatment. D ) CXCL12 transcript levels were higher in FIV-infected animals but were not reduced by ddI treatment. E ) MMP-2 transcripts were unregulated in FIV-infected animals but were suppressed with ddI treatment. F ) GFAP transcriptional activity was not affected by FIV infection or ddI treatment. * P

Techniques Used: Infection, Activity Assay

Suppression of neuronal autophagy and its regulation by antiretroviral therapy. ddI suppressed viral burden in the peripheral circulation, diminishing leukocyte activation and CNS entry with an ensuing reduction in MMP-2 production and its actions, thereby preventing the induction of CXCL12(5-12) with its adverse effects on neuronal autophagy and survival.
Figure Legend Snippet: Suppression of neuronal autophagy and its regulation by antiretroviral therapy. ddI suppressed viral burden in the peripheral circulation, diminishing leukocyte activation and CNS entry with an ensuing reduction in MMP-2 production and its actions, thereby preventing the induction of CXCL12(5-12) with its adverse effects on neuronal autophagy and survival.

Techniques Used: Activation Assay

33) Product Images from "CXCR3 activation by lentivirus infection suppresses neuronal autophagy: neuroprotective effects of antiretroviral therapy"

Article Title: CXCR3 activation by lentivirus infection suppresses neuronal autophagy: neuroprotective effects of antiretroviral therapy

Journal: The FASEB Journal

doi: 10.1096/fj.08-128819

Brain expression of CXCL12, MMP-2, and CXCR3 in FIV infection. A ) Immunoblotting showed increased full-length (1-67) and cleaved (5-67) CXCL12 in brains of FIV − compared with mock-infected animals. B ) CXCL12, CXCL12(5-67), MMP-2, and CXCR3 immunoreactivities were increased in brains of FIV + animals but were suppressed by concurrent ddI treatment. C ) MMP-2 immunoreactivity (IR) showed minimal induction in FIV + animals with and without ddI treatment. D , E ) CXCL12(1-67) ( D ) and CXCL12(5-67) ( E ) were induced by FIV infection, but these changes were reversed by ddI treatment. F ) CXCR3 showed minimal induction of both immunoreactive bands in FIV infection and was not affected by ddI treatment. * P
Figure Legend Snippet: Brain expression of CXCL12, MMP-2, and CXCR3 in FIV infection. A ) Immunoblotting showed increased full-length (1-67) and cleaved (5-67) CXCL12 in brains of FIV − compared with mock-infected animals. B ) CXCL12, CXCL12(5-67), MMP-2, and CXCR3 immunoreactivities were increased in brains of FIV + animals but were suppressed by concurrent ddI treatment. C ) MMP-2 immunoreactivity (IR) showed minimal induction in FIV + animals with and without ddI treatment. D , E ) CXCL12(1-67) ( D ) and CXCL12(5-67) ( E ) were induced by FIV infection, but these changes were reversed by ddI treatment. F ) CXCR3 showed minimal induction of both immunoreactive bands in FIV infection and was not affected by ddI treatment. * P

Techniques Used: Expressing, Infection

Neurobehavioral performance, viral burden, and neuroinflammation in FIV infection. A ) FIV-infected animals (FIV+) exhibit greater neurobehavioral deficits than mock-infected controls (FIV−), while ddI treatment reversed neurobehavioral deficits in FIV-infected animals (FIV+/ddI) and had no effects on mock-infected animals. B ) ddI treatment suppressed viral load in plasma of FIV-infected animals. C ) Glycoprotein, F4/80 , expressed in activated myeloid cells showed increased transcript abundance in FIV-infected animals, which was reversed by ddI treatment. D ) CXCL12 transcript levels were higher in FIV-infected animals but were not reduced by ddI treatment. E ) MMP-2 transcripts were unregulated in FIV-infected animals but were suppressed with ddI treatment. F ) GFAP transcriptional activity was not affected by FIV infection or ddI treatment. * P
Figure Legend Snippet: Neurobehavioral performance, viral burden, and neuroinflammation in FIV infection. A ) FIV-infected animals (FIV+) exhibit greater neurobehavioral deficits than mock-infected controls (FIV−), while ddI treatment reversed neurobehavioral deficits in FIV-infected animals (FIV+/ddI) and had no effects on mock-infected animals. B ) ddI treatment suppressed viral load in plasma of FIV-infected animals. C ) Glycoprotein, F4/80 , expressed in activated myeloid cells showed increased transcript abundance in FIV-infected animals, which was reversed by ddI treatment. D ) CXCL12 transcript levels were higher in FIV-infected animals but were not reduced by ddI treatment. E ) MMP-2 transcripts were unregulated in FIV-infected animals but were suppressed with ddI treatment. F ) GFAP transcriptional activity was not affected by FIV infection or ddI treatment. * P

Techniques Used: Infection, Activity Assay

Suppression of neuronal autophagy and its regulation by antiretroviral therapy. ddI suppressed viral burden in the peripheral circulation, diminishing leukocyte activation and CNS entry with an ensuing reduction in MMP-2 production and its actions, thereby preventing the induction of CXCL12(5-12) with its adverse effects on neuronal autophagy and survival.
Figure Legend Snippet: Suppression of neuronal autophagy and its regulation by antiretroviral therapy. ddI suppressed viral burden in the peripheral circulation, diminishing leukocyte activation and CNS entry with an ensuing reduction in MMP-2 production and its actions, thereby preventing the induction of CXCL12(5-12) with its adverse effects on neuronal autophagy and survival.

Techniques Used: Activation Assay

34) Product Images from "CXCR3 activation by lentivirus infection suppresses neuronal autophagy: neuroprotective effects of antiretroviral therapy"

Article Title: CXCR3 activation by lentivirus infection suppresses neuronal autophagy: neuroprotective effects of antiretroviral therapy

Journal: The FASEB Journal

doi: 10.1096/fj.08-128819

Brain expression of CXCL12, MMP-2, and CXCR3 in FIV infection. A ) Immunoblotting showed increased full-length (1-67) and cleaved (5-67) CXCL12 in brains of FIV − compared with mock-infected animals. B ) CXCL12, CXCL12(5-67), MMP-2, and CXCR3 immunoreactivities were increased in brains of FIV + animals but were suppressed by concurrent ddI treatment. C ) MMP-2 immunoreactivity (IR) showed minimal induction in FIV + animals with and without ddI treatment. D , E ) CXCL12(1-67) ( D ) and CXCL12(5-67) ( E ) were induced by FIV infection, but these changes were reversed by ddI treatment. F ) CXCR3 showed minimal induction of both immunoreactive bands in FIV infection and was not affected by ddI treatment. * P
Figure Legend Snippet: Brain expression of CXCL12, MMP-2, and CXCR3 in FIV infection. A ) Immunoblotting showed increased full-length (1-67) and cleaved (5-67) CXCL12 in brains of FIV − compared with mock-infected animals. B ) CXCL12, CXCL12(5-67), MMP-2, and CXCR3 immunoreactivities were increased in brains of FIV + animals but were suppressed by concurrent ddI treatment. C ) MMP-2 immunoreactivity (IR) showed minimal induction in FIV + animals with and without ddI treatment. D , E ) CXCL12(1-67) ( D ) and CXCL12(5-67) ( E ) were induced by FIV infection, but these changes were reversed by ddI treatment. F ) CXCR3 showed minimal induction of both immunoreactive bands in FIV infection and was not affected by ddI treatment. * P

Techniques Used: Expressing, Infection

Neurobehavioral performance, viral burden, and neuroinflammation in FIV infection. A ) FIV-infected animals (FIV+) exhibit greater neurobehavioral deficits than mock-infected controls (FIV−), while ddI treatment reversed neurobehavioral deficits in FIV-infected animals (FIV+/ddI) and had no effects on mock-infected animals. B ) ddI treatment suppressed viral load in plasma of FIV-infected animals. C ) Glycoprotein, F4/80 , expressed in activated myeloid cells showed increased transcript abundance in FIV-infected animals, which was reversed by ddI treatment. D ) CXCL12 transcript levels were higher in FIV-infected animals but were not reduced by ddI treatment. E ) MMP-2 transcripts were unregulated in FIV-infected animals but were suppressed with ddI treatment. F ) GFAP transcriptional activity was not affected by FIV infection or ddI treatment. * P
Figure Legend Snippet: Neurobehavioral performance, viral burden, and neuroinflammation in FIV infection. A ) FIV-infected animals (FIV+) exhibit greater neurobehavioral deficits than mock-infected controls (FIV−), while ddI treatment reversed neurobehavioral deficits in FIV-infected animals (FIV+/ddI) and had no effects on mock-infected animals. B ) ddI treatment suppressed viral load in plasma of FIV-infected animals. C ) Glycoprotein, F4/80 , expressed in activated myeloid cells showed increased transcript abundance in FIV-infected animals, which was reversed by ddI treatment. D ) CXCL12 transcript levels were higher in FIV-infected animals but were not reduced by ddI treatment. E ) MMP-2 transcripts were unregulated in FIV-infected animals but were suppressed with ddI treatment. F ) GFAP transcriptional activity was not affected by FIV infection or ddI treatment. * P

Techniques Used: Infection, Activity Assay

Suppression of neuronal autophagy and its regulation by antiretroviral therapy. ddI suppressed viral burden in the peripheral circulation, diminishing leukocyte activation and CNS entry with an ensuing reduction in MMP-2 production and its actions, thereby preventing the induction of CXCL12(5-12) with its adverse effects on neuronal autophagy and survival.
Figure Legend Snippet: Suppression of neuronal autophagy and its regulation by antiretroviral therapy. ddI suppressed viral burden in the peripheral circulation, diminishing leukocyte activation and CNS entry with an ensuing reduction in MMP-2 production and its actions, thereby preventing the induction of CXCL12(5-12) with its adverse effects on neuronal autophagy and survival.

Techniques Used: Activation Assay

35) Product Images from "Association of MMP-2, RB and PAI-1 with decreased recurrence-free survival and overall survival in bladder cancer patients"

Article Title: Association of MMP-2, RB and PAI-1 with decreased recurrence-free survival and overall survival in bladder cancer patients

Journal: Oncotarget

doi: 10.18632/oncotarget.20686

Progression-free survival analysis of 939 patients with bladder cancer Progression-free survival according to immunostaining status of ANG, MMP-2, p53, RB and PAI-1 in NMIBC (A) and MIBC (B) .
Figure Legend Snippet: Progression-free survival analysis of 939 patients with bladder cancer Progression-free survival according to immunostaining status of ANG, MMP-2, p53, RB and PAI-1 in NMIBC (A) and MIBC (B) .

Techniques Used: Immunostaining

Representative expression status for ANG, MMP-2, p53, RB and PAI-1 in a high-grade non-muscle invasive bladder cancer Upper row represents high intensity while lower row represents low intensity associated with each target. All images were captured at 400× magnification.
Figure Legend Snippet: Representative expression status for ANG, MMP-2, p53, RB and PAI-1 in a high-grade non-muscle invasive bladder cancer Upper row represents high intensity while lower row represents low intensity associated with each target. All images were captured at 400× magnification.

Techniques Used: Expressing

Overall survival analysis of 939 patients with bladder cancer Overall survival according to immunostaining status of ANG, MMP-2, p53, RB and PAI-1 in NMIBC (A) and MIBC (B) .
Figure Legend Snippet: Overall survival analysis of 939 patients with bladder cancer Overall survival according to immunostaining status of ANG, MMP-2, p53, RB and PAI-1 in NMIBC (A) and MIBC (B) .

Techniques Used: Immunostaining

36) Product Images from "Anti-angiogenic therapy increases intratumoral adenovirus distribution by inducing collagen degradation"

Article Title: Anti-angiogenic therapy increases intratumoral adenovirus distribution by inducing collagen degradation

Journal: Gene therapy

doi: 10.1038/gt.2012.42

VEGF trapping up-regulates MMP-2 expression in glioma cell lines. (A) Western blot of U87, U118, A172 and U251 glioma cells after treatment with VEGF-Ab for 5 days. Before collection, cell secretion was blocked for 6 hours with Golgi-Plug. Human β-Actin was used as a protein loading control. (B) Quantitative real-time PCR for MMP-2 mRNA levels in glioma cell lines was evaluated using the Δ C T method. Before plotted in the graph, the MMP-2 expression for each cell line was normalized to their Ig-Control treated condition, to which was given an arbitrary value of 1 (dotted, thin black line: Control). The relative expression levels of MMP-2 for all four cell lines upon VEGF-Ab treatment were then presented as fold of their Control treated cells at the indicated time points. (C) Immunocytochemistry (ICC) of glioma cell lines for MMP-2 and collagen IV after treatment with VEGF-Ab. Cells treated in vitro with VEGF-Ab for 5 days were blocked with Golgi-Plug for 6 hours before fixation for ICC. Nuclei are stained blue with DAPI; collagen IV is presented in yellow and MMP-2 in red. Bars equal 50 μ m .
Figure Legend Snippet: VEGF trapping up-regulates MMP-2 expression in glioma cell lines. (A) Western blot of U87, U118, A172 and U251 glioma cells after treatment with VEGF-Ab for 5 days. Before collection, cell secretion was blocked for 6 hours with Golgi-Plug. Human β-Actin was used as a protein loading control. (B) Quantitative real-time PCR for MMP-2 mRNA levels in glioma cell lines was evaluated using the Δ C T method. Before plotted in the graph, the MMP-2 expression for each cell line was normalized to their Ig-Control treated condition, to which was given an arbitrary value of 1 (dotted, thin black line: Control). The relative expression levels of MMP-2 for all four cell lines upon VEGF-Ab treatment were then presented as fold of their Control treated cells at the indicated time points. (C) Immunocytochemistry (ICC) of glioma cell lines for MMP-2 and collagen IV after treatment with VEGF-Ab. Cells treated in vitro with VEGF-Ab for 5 days were blocked with Golgi-Plug for 6 hours before fixation for ICC. Nuclei are stained blue with DAPI; collagen IV is presented in yellow and MMP-2 in red. Bars equal 50 μ m .

Techniques Used: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Immunocytochemistry, In Vitro, Staining

Anti-angiogenic treatment alters the extracellular matrix (ECM) architecture of human glioma xenograft in nude mice. (A) Immunohistochemistry staining for MMP-2, Collagen IV, CD31 and Laminin. (B–E) Quantification of staining intensity was done through a computer based scoring for each of the corresponding IHC slides (n = 5 animals for each group) and mean values ± standard error of measurement (SEM) are presented in bar diagrams. Bars equal 50 μ m; *p
Figure Legend Snippet: Anti-angiogenic treatment alters the extracellular matrix (ECM) architecture of human glioma xenograft in nude mice. (A) Immunohistochemistry staining for MMP-2, Collagen IV, CD31 and Laminin. (B–E) Quantification of staining intensity was done through a computer based scoring for each of the corresponding IHC slides (n = 5 animals for each group) and mean values ± standard error of measurement (SEM) are presented in bar diagrams. Bars equal 50 μ m; *p

Techniques Used: Mouse Assay, Immunohistochemistry, Staining

Adenovirus infection does not increase MMP-2 expression. U251 xenografts sections (A) were stained for MMP-2 with HRP and counterstained with hematoxylin; then scanned for intensity of HRP staining, presented as percent of areas with similar intensity (B). Bars represent mean intensity values of all xenografts. ns, not significant ; *** p
Figure Legend Snippet: Adenovirus infection does not increase MMP-2 expression. U251 xenografts sections (A) were stained for MMP-2 with HRP and counterstained with hematoxylin; then scanned for intensity of HRP staining, presented as percent of areas with similar intensity (B). Bars represent mean intensity values of all xenografts. ns, not significant ; *** p

Techniques Used: Infection, Expressing, Staining

37) Product Images from "CXCR3 activation by lentivirus infection suppresses neuronal autophagy: neuroprotective effects of antiretroviral therapy"

Article Title: CXCR3 activation by lentivirus infection suppresses neuronal autophagy: neuroprotective effects of antiretroviral therapy

Journal: The FASEB Journal

doi: 10.1096/fj.08-128819

Brain expression of CXCL12, MMP-2, and CXCR3 in FIV infection. A ) Immunoblotting showed increased full-length (1-67) and cleaved (5-67) CXCL12 in brains of FIV − compared with mock-infected animals. B ) CXCL12, CXCL12(5-67), MMP-2, and CXCR3 immunoreactivities were increased in brains of FIV + animals but were suppressed by concurrent ddI treatment. C ) MMP-2 immunoreactivity (IR) showed minimal induction in FIV + animals with and without ddI treatment. D , E ) CXCL12(1-67) ( D ) and CXCL12(5-67) ( E ) were induced by FIV infection, but these changes were reversed by ddI treatment. F ) CXCR3 showed minimal induction of both immunoreactive bands in FIV infection and was not affected by ddI treatment. * P
Figure Legend Snippet: Brain expression of CXCL12, MMP-2, and CXCR3 in FIV infection. A ) Immunoblotting showed increased full-length (1-67) and cleaved (5-67) CXCL12 in brains of FIV − compared with mock-infected animals. B ) CXCL12, CXCL12(5-67), MMP-2, and CXCR3 immunoreactivities were increased in brains of FIV + animals but were suppressed by concurrent ddI treatment. C ) MMP-2 immunoreactivity (IR) showed minimal induction in FIV + animals with and without ddI treatment. D , E ) CXCL12(1-67) ( D ) and CXCL12(5-67) ( E ) were induced by FIV infection, but these changes were reversed by ddI treatment. F ) CXCR3 showed minimal induction of both immunoreactive bands in FIV infection and was not affected by ddI treatment. * P

Techniques Used: Expressing, Infection

Neurobehavioral performance, viral burden, and neuroinflammation in FIV infection. A ) FIV-infected animals (FIV+) exhibit greater neurobehavioral deficits than mock-infected controls (FIV−), while ddI treatment reversed neurobehavioral deficits in FIV-infected animals (FIV+/ddI) and had no effects on mock-infected animals. B ) ddI treatment suppressed viral load in plasma of FIV-infected animals. C ) Glycoprotein, F4/80 , expressed in activated myeloid cells showed increased transcript abundance in FIV-infected animals, which was reversed by ddI treatment. D ) CXCL12 transcript levels were higher in FIV-infected animals but were not reduced by ddI treatment. E ) MMP-2 transcripts were unregulated in FIV-infected animals but were suppressed with ddI treatment. F ) GFAP transcriptional activity was not affected by FIV infection or ddI treatment. * P
Figure Legend Snippet: Neurobehavioral performance, viral burden, and neuroinflammation in FIV infection. A ) FIV-infected animals (FIV+) exhibit greater neurobehavioral deficits than mock-infected controls (FIV−), while ddI treatment reversed neurobehavioral deficits in FIV-infected animals (FIV+/ddI) and had no effects on mock-infected animals. B ) ddI treatment suppressed viral load in plasma of FIV-infected animals. C ) Glycoprotein, F4/80 , expressed in activated myeloid cells showed increased transcript abundance in FIV-infected animals, which was reversed by ddI treatment. D ) CXCL12 transcript levels were higher in FIV-infected animals but were not reduced by ddI treatment. E ) MMP-2 transcripts were unregulated in FIV-infected animals but were suppressed with ddI treatment. F ) GFAP transcriptional activity was not affected by FIV infection or ddI treatment. * P

Techniques Used: Infection, Activity Assay

Suppression of neuronal autophagy and its regulation by antiretroviral therapy. ddI suppressed viral burden in the peripheral circulation, diminishing leukocyte activation and CNS entry with an ensuing reduction in MMP-2 production and its actions, thereby preventing the induction of CXCL12(5-12) with its adverse effects on neuronal autophagy and survival.
Figure Legend Snippet: Suppression of neuronal autophagy and its regulation by antiretroviral therapy. ddI suppressed viral burden in the peripheral circulation, diminishing leukocyte activation and CNS entry with an ensuing reduction in MMP-2 production and its actions, thereby preventing the induction of CXCL12(5-12) with its adverse effects on neuronal autophagy and survival.

Techniques Used: Activation Assay

38) Product Images from "Anti-angiogenic therapy increases intratumoral adenovirus distribution by inducing collagen degradation"

Article Title: Anti-angiogenic therapy increases intratumoral adenovirus distribution by inducing collagen degradation

Journal: Gene therapy

doi: 10.1038/gt.2012.42

VEGF trapping up-regulates MMP-2 expression in glioma cell lines. (A) Western blot of U87, U118, A172 and U251 glioma cells after treatment with VEGF-Ab for 5 days. Before collection, cell secretion was blocked for 6 hours with Golgi-Plug. Human β-Actin was used as a protein loading control. (B) Quantitative real-time PCR for MMP-2 mRNA levels in glioma cell lines was evaluated using the Δ C T method. Before plotted in the graph, the MMP-2 expression for each cell line was normalized to their Ig-Control treated condition, to which was given an arbitrary value of 1 (dotted, thin black line: Control). The relative expression levels of MMP-2 for all four cell lines upon VEGF-Ab treatment were then presented as fold of their Control treated cells at the indicated time points. (C) Immunocytochemistry (ICC) of glioma cell lines for MMP-2 and collagen IV after treatment with VEGF-Ab. Cells treated in vitro with VEGF-Ab for 5 days were blocked with Golgi-Plug for 6 hours before fixation for ICC. Nuclei are stained blue with DAPI; collagen IV is presented in yellow and MMP-2 in red. Bars equal 50 μ m .
Figure Legend Snippet: VEGF trapping up-regulates MMP-2 expression in glioma cell lines. (A) Western blot of U87, U118, A172 and U251 glioma cells after treatment with VEGF-Ab for 5 days. Before collection, cell secretion was blocked for 6 hours with Golgi-Plug. Human β-Actin was used as a protein loading control. (B) Quantitative real-time PCR for MMP-2 mRNA levels in glioma cell lines was evaluated using the Δ C T method. Before plotted in the graph, the MMP-2 expression for each cell line was normalized to their Ig-Control treated condition, to which was given an arbitrary value of 1 (dotted, thin black line: Control). The relative expression levels of MMP-2 for all four cell lines upon VEGF-Ab treatment were then presented as fold of their Control treated cells at the indicated time points. (C) Immunocytochemistry (ICC) of glioma cell lines for MMP-2 and collagen IV after treatment with VEGF-Ab. Cells treated in vitro with VEGF-Ab for 5 days were blocked with Golgi-Plug for 6 hours before fixation for ICC. Nuclei are stained blue with DAPI; collagen IV is presented in yellow and MMP-2 in red. Bars equal 50 μ m .

Techniques Used: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Immunocytochemistry, In Vitro, Staining

Anti-angiogenic treatment alters the extracellular matrix (ECM) architecture of human glioma xenograft in nude mice. (A) Immunohistochemistry staining for MMP-2, Collagen IV, CD31 and Laminin. (B–E) Quantification of staining intensity was done through a computer based scoring for each of the corresponding IHC slides (n = 5 animals for each group) and mean values ± standard error of measurement (SEM) are presented in bar diagrams. Bars equal 50 μ m; *p
Figure Legend Snippet: Anti-angiogenic treatment alters the extracellular matrix (ECM) architecture of human glioma xenograft in nude mice. (A) Immunohistochemistry staining for MMP-2, Collagen IV, CD31 and Laminin. (B–E) Quantification of staining intensity was done through a computer based scoring for each of the corresponding IHC slides (n = 5 animals for each group) and mean values ± standard error of measurement (SEM) are presented in bar diagrams. Bars equal 50 μ m; *p

Techniques Used: Mouse Assay, Immunohistochemistry, Staining

Adenovirus infection does not increase MMP-2 expression. U251 xenografts sections (A) were stained for MMP-2 with HRP and counterstained with hematoxylin; then scanned for intensity of HRP staining, presented as percent of areas with similar intensity (B). Bars represent mean intensity values of all xenografts. ns, not significant ; *** p
Figure Legend Snippet: Adenovirus infection does not increase MMP-2 expression. U251 xenografts sections (A) were stained for MMP-2 with HRP and counterstained with hematoxylin; then scanned for intensity of HRP staining, presented as percent of areas with similar intensity (B). Bars represent mean intensity values of all xenografts. ns, not significant ; *** p

Techniques Used: Infection, Expressing, Staining

39) Product Images from "Anti-angiogenic therapy increases intratumoral adenovirus distribution by inducing collagen degradation"

Article Title: Anti-angiogenic therapy increases intratumoral adenovirus distribution by inducing collagen degradation

Journal: Gene therapy

doi: 10.1038/gt.2012.42

VEGF trapping up-regulates MMP-2 expression in glioma cell lines. (A) Western blot of U87, U118, A172 and U251 glioma cells after treatment with VEGF-Ab for 5 days. Before collection, cell secretion was blocked for 6 hours with Golgi-Plug. Human β-Actin was used as a protein loading control. (B) Quantitative real-time PCR for MMP-2 mRNA levels in glioma cell lines was evaluated using the Δ C T method. Before plotted in the graph, the MMP-2 expression for each cell line was normalized to their Ig-Control treated condition, to which was given an arbitrary value of 1 (dotted, thin black line: Control). The relative expression levels of MMP-2 for all four cell lines upon VEGF-Ab treatment were then presented as fold of their Control treated cells at the indicated time points. (C) Immunocytochemistry (ICC) of glioma cell lines for MMP-2 and collagen IV after treatment with VEGF-Ab. Cells treated in vitro with VEGF-Ab for 5 days were blocked with Golgi-Plug for 6 hours before fixation for ICC. Nuclei are stained blue with DAPI; collagen IV is presented in yellow and MMP-2 in red. Bars equal 50 μ m .
Figure Legend Snippet: VEGF trapping up-regulates MMP-2 expression in glioma cell lines. (A) Western blot of U87, U118, A172 and U251 glioma cells after treatment with VEGF-Ab for 5 days. Before collection, cell secretion was blocked for 6 hours with Golgi-Plug. Human β-Actin was used as a protein loading control. (B) Quantitative real-time PCR for MMP-2 mRNA levels in glioma cell lines was evaluated using the Δ C T method. Before plotted in the graph, the MMP-2 expression for each cell line was normalized to their Ig-Control treated condition, to which was given an arbitrary value of 1 (dotted, thin black line: Control). The relative expression levels of MMP-2 for all four cell lines upon VEGF-Ab treatment were then presented as fold of their Control treated cells at the indicated time points. (C) Immunocytochemistry (ICC) of glioma cell lines for MMP-2 and collagen IV after treatment with VEGF-Ab. Cells treated in vitro with VEGF-Ab for 5 days were blocked with Golgi-Plug for 6 hours before fixation for ICC. Nuclei are stained blue with DAPI; collagen IV is presented in yellow and MMP-2 in red. Bars equal 50 μ m .

Techniques Used: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Immunocytochemistry, In Vitro, Staining

Anti-angiogenic treatment alters the extracellular matrix (ECM) architecture of human glioma xenograft in nude mice. (A) Immunohistochemistry staining for MMP-2, Collagen IV, CD31 and Laminin. (B–E) Quantification of staining intensity was done through a computer based scoring for each of the corresponding IHC slides (n = 5 animals for each group) and mean values ± standard error of measurement (SEM) are presented in bar diagrams. Bars equal 50 μ m; *p
Figure Legend Snippet: Anti-angiogenic treatment alters the extracellular matrix (ECM) architecture of human glioma xenograft in nude mice. (A) Immunohistochemistry staining for MMP-2, Collagen IV, CD31 and Laminin. (B–E) Quantification of staining intensity was done through a computer based scoring for each of the corresponding IHC slides (n = 5 animals for each group) and mean values ± standard error of measurement (SEM) are presented in bar diagrams. Bars equal 50 μ m; *p

Techniques Used: Mouse Assay, Immunohistochemistry, Staining

Adenovirus infection does not increase MMP-2 expression. U251 xenografts sections (A) were stained for MMP-2 with HRP and counterstained with hematoxylin; then scanned for intensity of HRP staining, presented as percent of areas with similar intensity (B). Bars represent mean intensity values of all xenografts. ns, not significant ; *** p
Figure Legend Snippet: Adenovirus infection does not increase MMP-2 expression. U251 xenografts sections (A) were stained for MMP-2 with HRP and counterstained with hematoxylin; then scanned for intensity of HRP staining, presented as percent of areas with similar intensity (B). Bars represent mean intensity values of all xenografts. ns, not significant ; *** p

Techniques Used: Infection, Expressing, Staining

40) Product Images from "Curcumin derivative WZ35 inhibits tumor cell growth via ROS-YAP-JNK signaling pathway in breast cancer"

Article Title: Curcumin derivative WZ35 inhibits tumor cell growth via ROS-YAP-JNK signaling pathway in breast cancer

Journal: Journal of Experimental & Clinical Cancer Research : CR

doi: 10.1186/s13046-019-1424-4

WZ35 inhibits breast cancer cells proliferation, migration, invasion and induced apoptotic cell death and cell cycle arrest. a The effects of WZ35 and curcumin on the proliferation of breast cancer cells. b Induction of cycle arrest in MDA-MB-231 cells was detected by Flow cytometry after treatment with curcumin (10 μg/mL) and WZ35 (10 μg/mL) for 24 h. WZ53 increased the proportion of cells in the G2/M phase. c Expression of cell cycle relative proteins Cyclin B1, and p21 were determined by western blot after treatment with WZ35 (10 μg/mL) or curcumin (10 μg/mL) for 24 h. GAPDH was used as internal control. d Induction of apoptosis in MDA-MB-231 cells was determined by flow cytometry after treatment with WZ35 (10 μg/mL) and curcumin (10 μg/mL) for 24 h. Similar results were obtained in three independent experiments. e , h Transwell assay was performed to evaluate the effects of WZ35 and curcumin on Hs578T ( e , f ) and MDA-MB-231 ( g , h ) cells migration and invasion. Cell migration and invasion images were presented and migrated cells were quantified. i EMT biomakers N-cadherin, E-cadherin, MMP-2 and MMP-9 were determined by western blot. Data are presented as mean ± SD, * p
Figure Legend Snippet: WZ35 inhibits breast cancer cells proliferation, migration, invasion and induced apoptotic cell death and cell cycle arrest. a The effects of WZ35 and curcumin on the proliferation of breast cancer cells. b Induction of cycle arrest in MDA-MB-231 cells was detected by Flow cytometry after treatment with curcumin (10 μg/mL) and WZ35 (10 μg/mL) for 24 h. WZ53 increased the proportion of cells in the G2/M phase. c Expression of cell cycle relative proteins Cyclin B1, and p21 were determined by western blot after treatment with WZ35 (10 μg/mL) or curcumin (10 μg/mL) for 24 h. GAPDH was used as internal control. d Induction of apoptosis in MDA-MB-231 cells was determined by flow cytometry after treatment with WZ35 (10 μg/mL) and curcumin (10 μg/mL) for 24 h. Similar results were obtained in three independent experiments. e , h Transwell assay was performed to evaluate the effects of WZ35 and curcumin on Hs578T ( e , f ) and MDA-MB-231 ( g , h ) cells migration and invasion. Cell migration and invasion images were presented and migrated cells were quantified. i EMT biomakers N-cadherin, E-cadherin, MMP-2 and MMP-9 were determined by western blot. Data are presented as mean ± SD, * p

Techniques Used: Migration, Multiple Displacement Amplification, Flow Cytometry, Cytometry, Expressing, Western Blot, Transwell Assay

WZ35 inhibits MDA-MB-231 xenograft tumor growth and metastasis in vivo. a A growth curve analysis of the tumor growth in curcumin, WZ35 or Castor oil treated groups. b Expression of ki67 in tumor tissues was determined by immunohistochemistry. c Mice body weight was measured during the 17-day treatment of WZ35 and curcumin. d Representative microscopic images of lung metastatic lesions at 21 days after treatment of mice with curcumin or WZ35 (0.2 mL, 25 mg/kg for each). e The number of lung metastatic tumors was calculated. f Immunofluorescence were used to test the expression level of E-cadherin, N-cadherin, MMP-2 and MMP-9 from curcumin, WZ35, Vehicle and control group. Data are presented as mean ± SD, *** p
Figure Legend Snippet: WZ35 inhibits MDA-MB-231 xenograft tumor growth and metastasis in vivo. a A growth curve analysis of the tumor growth in curcumin, WZ35 or Castor oil treated groups. b Expression of ki67 in tumor tissues was determined by immunohistochemistry. c Mice body weight was measured during the 17-day treatment of WZ35 and curcumin. d Representative microscopic images of lung metastatic lesions at 21 days after treatment of mice with curcumin or WZ35 (0.2 mL, 25 mg/kg for each). e The number of lung metastatic tumors was calculated. f Immunofluorescence were used to test the expression level of E-cadherin, N-cadherin, MMP-2 and MMP-9 from curcumin, WZ35, Vehicle and control group. Data are presented as mean ± SD, *** p

Techniques Used: Multiple Displacement Amplification, In Vivo, Expressing, Immunohistochemistry, Mouse Assay, Immunofluorescence

41) Product Images from "Inhibition of Radiation and Temozolomide-Induced Invadopodia Activity in Glioma Cells Using FDA-Approved Drugs"

Article Title: Inhibition of Radiation and Temozolomide-Induced Invadopodia Activity in Glioma Cells Using FDA-Approved Drugs

Journal: Translational Oncology

doi: 10.1016/j.tranon.2018.08.012

GBM cell lines express invadopodia regulators, form functional invadopodia and secrete MMP-2. (A) LN229, U87MG, U87MG-EGFRvIII, MU35 and MU41 GBM cells were plated on a thin film of cross-linked FITC-gelatin for 24 hours to detect the presence of invadopodia-mediated FITC-gelatin degradation. White scale bars represent 20 μm. Mean FITC-labeled gelatin degrading activity of GBM cells. Degraded areas of FITC-labeled gelatin are evident as black areas devoid of FITC-labeled gelatin (green). DAPI staining of the nucleus is shown in blue, and rhodamine phalloidin (red) was used to stain for actin filaments and actin puncta (invadopodia). (B) Graph depicting the basal invadopodia-mediated FITC-gelatin degradation activity of the GBM cell lines in (A). Experiment repeated two times. (C) Immunoblot analysis of various regulators of invadopodia formation or activity in the listed GBM cell lines. The experiment was repeated twice, and representative images are shown. (D) Gelatin zymogram analysis showing MMP-2 activity at 24 hours after incubation of GBM cells in serum-free Optimem. The experiment was repeated twice, and a representative image is shown.
Figure Legend Snippet: GBM cell lines express invadopodia regulators, form functional invadopodia and secrete MMP-2. (A) LN229, U87MG, U87MG-EGFRvIII, MU35 and MU41 GBM cells were plated on a thin film of cross-linked FITC-gelatin for 24 hours to detect the presence of invadopodia-mediated FITC-gelatin degradation. White scale bars represent 20 μm. Mean FITC-labeled gelatin degrading activity of GBM cells. Degraded areas of FITC-labeled gelatin are evident as black areas devoid of FITC-labeled gelatin (green). DAPI staining of the nucleus is shown in blue, and rhodamine phalloidin (red) was used to stain for actin filaments and actin puncta (invadopodia). (B) Graph depicting the basal invadopodia-mediated FITC-gelatin degradation activity of the GBM cell lines in (A). Experiment repeated two times. (C) Immunoblot analysis of various regulators of invadopodia formation or activity in the listed GBM cell lines. The experiment was repeated twice, and representative images are shown. (D) Gelatin zymogram analysis showing MMP-2 activity at 24 hours after incubation of GBM cells in serum-free Optimem. The experiment was repeated twice, and a representative image is shown.

Techniques Used: Functional Assay, Labeling, Activity Assay, Staining, Incubation

MMP-2 secretion and invadopodia-mediated FITC-gelatin degradation are enhanced by radiation and TMZ. Gelatin-based zymogram (A) and FITC-gelatin degradation (B) analyses of LN229 GBM cells treated with RT/TMZ (2 Gy/200 μM). Zymographic analysis of serum-free Optimem was performed 24 hours posttreatment. For FITC-gelatin degradation assays, cells were trypsinized 24 hours posttreatment and seeded on FITC-labeled gelatin for 24 hours, after which the cells were subsequently fixed and stained for actin filaments with rhodamine-phalloidin (red) and DAPI nuclear staining (blue). Degraded areas of FITC-labeled gelatin are evident as black areas devoid of FITC-labeled gelatin (green). The mean FITC-labeled gelatin-degrading activity (relative to the RT+TMZ treated cells) was determined (B). * P
Figure Legend Snippet: MMP-2 secretion and invadopodia-mediated FITC-gelatin degradation are enhanced by radiation and TMZ. Gelatin-based zymogram (A) and FITC-gelatin degradation (B) analyses of LN229 GBM cells treated with RT/TMZ (2 Gy/200 μM). Zymographic analysis of serum-free Optimem was performed 24 hours posttreatment. For FITC-gelatin degradation assays, cells were trypsinized 24 hours posttreatment and seeded on FITC-labeled gelatin for 24 hours, after which the cells were subsequently fixed and stained for actin filaments with rhodamine-phalloidin (red) and DAPI nuclear staining (blue). Degraded areas of FITC-labeled gelatin are evident as black areas devoid of FITC-labeled gelatin (green). The mean FITC-labeled gelatin-degrading activity (relative to the RT+TMZ treated cells) was determined (B). * P

Techniques Used: Labeling, Staining, Activity Assay

Vinorelbine tartrate and paclitaxel can reduce MMP-2 secretion in GBM cells. (A) Gelatin-based zymogram analysis of serum-free Optimem conditioned medium showing MMP-2 secretion activity of MU41 and LN229 GBM cells treated with 0.01 μM paclitaxel (PAC), vinorelbine tartrate (VT), fludarabine phosphate (FP), or dasatinib (DAS) [untreated control (CON)] for 24 hours. The experiment was repeated twice, and a representative image is shown. (B) Densitometric analysis of zymograms.
Figure Legend Snippet: Vinorelbine tartrate and paclitaxel can reduce MMP-2 secretion in GBM cells. (A) Gelatin-based zymogram analysis of serum-free Optimem conditioned medium showing MMP-2 secretion activity of MU41 and LN229 GBM cells treated with 0.01 μM paclitaxel (PAC), vinorelbine tartrate (VT), fludarabine phosphate (FP), or dasatinib (DAS) [untreated control (CON)] for 24 hours. The experiment was repeated twice, and a representative image is shown. (B) Densitometric analysis of zymograms.

Techniques Used: Activity Assay

42) Product Images from "Inhibition of Radiation and Temozolomide-Induced Invadopodia Activity in Glioma Cells Using FDA-Approved Drugs"

Article Title: Inhibition of Radiation and Temozolomide-Induced Invadopodia Activity in Glioma Cells Using FDA-Approved Drugs

Journal: Translational Oncology

doi: 10.1016/j.tranon.2018.08.012

GBM cell lines express invadopodia regulators, form functional invadopodia and secrete MMP-2. (A) LN229, U87MG, U87MG-EGFRvIII, MU35 and MU41 GBM cells were plated on a thin film of cross-linked FITC-gelatin for 24 hours to detect the presence of invadopodia-mediated FITC-gelatin degradation. White scale bars represent 20 μm. Mean FITC-labeled gelatin degrading activity of GBM cells. Degraded areas of FITC-labeled gelatin are evident as black areas devoid of FITC-labeled gelatin (green). DAPI staining of the nucleus is shown in blue, and rhodamine phalloidin (red) was used to stain for actin filaments and actin puncta (invadopodia). (B) Graph depicting the basal invadopodia-mediated FITC-gelatin degradation activity of the GBM cell lines in (A). Experiment repeated two times. (C) Immunoblot analysis of various regulators of invadopodia formation or activity in the listed GBM cell lines. The experiment was repeated twice, and representative images are shown. (D) Gelatin zymogram analysis showing MMP-2 activity at 24 hours after incubation of GBM cells in serum-free Optimem. The experiment was repeated twice, and a representative image is shown.
Figure Legend Snippet: GBM cell lines express invadopodia regulators, form functional invadopodia and secrete MMP-2. (A) LN229, U87MG, U87MG-EGFRvIII, MU35 and MU41 GBM cells were plated on a thin film of cross-linked FITC-gelatin for 24 hours to detect the presence of invadopodia-mediated FITC-gelatin degradation. White scale bars represent 20 μm. Mean FITC-labeled gelatin degrading activity of GBM cells. Degraded areas of FITC-labeled gelatin are evident as black areas devoid of FITC-labeled gelatin (green). DAPI staining of the nucleus is shown in blue, and rhodamine phalloidin (red) was used to stain for actin filaments and actin puncta (invadopodia). (B) Graph depicting the basal invadopodia-mediated FITC-gelatin degradation activity of the GBM cell lines in (A). Experiment repeated two times. (C) Immunoblot analysis of various regulators of invadopodia formation or activity in the listed GBM cell lines. The experiment was repeated twice, and representative images are shown. (D) Gelatin zymogram analysis showing MMP-2 activity at 24 hours after incubation of GBM cells in serum-free Optimem. The experiment was repeated twice, and a representative image is shown.

Techniques Used: Functional Assay, Labeling, Activity Assay, Staining, Incubation

MMP-2 secretion and invadopodia-mediated FITC-gelatin degradation are enhanced by radiation and TMZ. Gelatin-based zymogram (A) and FITC-gelatin degradation (B) analyses of LN229 GBM cells treated with RT/TMZ (2 Gy/200 μM). Zymographic analysis of serum-free Optimem was performed 24 hours posttreatment. For FITC-gelatin degradation assays, cells were trypsinized 24 hours posttreatment and seeded on FITC-labeled gelatin for 24 hours, after which the cells were subsequently fixed and stained for actin filaments with rhodamine-phalloidin (red) and DAPI nuclear staining (blue). Degraded areas of FITC-labeled gelatin are evident as black areas devoid of FITC-labeled gelatin (green). The mean FITC-labeled gelatin-degrading activity (relative to the RT+TMZ treated cells) was determined (B). * P
Figure Legend Snippet: MMP-2 secretion and invadopodia-mediated FITC-gelatin degradation are enhanced by radiation and TMZ. Gelatin-based zymogram (A) and FITC-gelatin degradation (B) analyses of LN229 GBM cells treated with RT/TMZ (2 Gy/200 μM). Zymographic analysis of serum-free Optimem was performed 24 hours posttreatment. For FITC-gelatin degradation assays, cells were trypsinized 24 hours posttreatment and seeded on FITC-labeled gelatin for 24 hours, after which the cells were subsequently fixed and stained for actin filaments with rhodamine-phalloidin (red) and DAPI nuclear staining (blue). Degraded areas of FITC-labeled gelatin are evident as black areas devoid of FITC-labeled gelatin (green). The mean FITC-labeled gelatin-degrading activity (relative to the RT+TMZ treated cells) was determined (B). * P

Techniques Used: Labeling, Staining, Activity Assay

Vinorelbine tartrate and paclitaxel can reduce MMP-2 secretion in GBM cells. (A) Gelatin-based zymogram analysis of serum-free Optimem conditioned medium showing MMP-2 secretion activity of MU41 and LN229 GBM cells treated with 0.01 μM paclitaxel (PAC), vinorelbine tartrate (VT), fludarabine phosphate (FP), or dasatinib (DAS) [untreated control (CON)] for 24 hours. The experiment was repeated twice, and a representative image is shown. (B) Densitometric analysis of zymograms.
Figure Legend Snippet: Vinorelbine tartrate and paclitaxel can reduce MMP-2 secretion in GBM cells. (A) Gelatin-based zymogram analysis of serum-free Optimem conditioned medium showing MMP-2 secretion activity of MU41 and LN229 GBM cells treated with 0.01 μM paclitaxel (PAC), vinorelbine tartrate (VT), fludarabine phosphate (FP), or dasatinib (DAS) [untreated control (CON)] for 24 hours. The experiment was repeated twice, and a representative image is shown. (B) Densitometric analysis of zymograms.

Techniques Used: Activity Assay

43) Product Images from "?-Mangostin suppresses lipopolysaccharide-induced invasion by inhibiting matrix metalloproteinase-2/9 and increasing E-cadherin expression through extracellular signal-regulated kinase signaling in pancreatic cancer cells"

Article Title: ?-Mangostin suppresses lipopolysaccharide-induced invasion by inhibiting matrix metalloproteinase-2/9 and increasing E-cadherin expression through extracellular signal-regulated kinase signaling in pancreatic cancer cells

Journal: Oncology Letters

doi: 10.3892/ol.2013.1290

α-mangostin prevented the LPS-induced decrease in the expression of E-cadherin mRNA and the increase in the expression of MMP-9 and MMP-2 mRNA. (A) The mRNA expression levels of E-cadherin, MMP-9 and MMP-2 in BxPC-3 and MIAPaCa-2 cells were determined by RT-PCR. (B) Quantification of the mRNA levels. Data from at least three independent experiments with duplicate determinations are expressed as the mean ± SEM. * P
Figure Legend Snippet: α-mangostin prevented the LPS-induced decrease in the expression of E-cadherin mRNA and the increase in the expression of MMP-9 and MMP-2 mRNA. (A) The mRNA expression levels of E-cadherin, MMP-9 and MMP-2 in BxPC-3 and MIAPaCa-2 cells were determined by RT-PCR. (B) Quantification of the mRNA levels. Data from at least three independent experiments with duplicate determinations are expressed as the mean ± SEM. * P

Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

α-mangostin prevented the LPS-induced decrease in the expression of E-cadherin protein and LPS-induced increase in the expression of MMP-9 and MMP-2 protein. (A) Protein expression levels of E-cadherin, MMP-9, and MMP-2 in BxPC-3 and MIAPaCa-2 cells were determined via western blotting. (B) Quantification of the protein levels. Data from at least three independent experiments with duplicate determinations are expressed as the mean ± SEM. * P
Figure Legend Snippet: α-mangostin prevented the LPS-induced decrease in the expression of E-cadherin protein and LPS-induced increase in the expression of MMP-9 and MMP-2 protein. (A) Protein expression levels of E-cadherin, MMP-9, and MMP-2 in BxPC-3 and MIAPaCa-2 cells were determined via western blotting. (B) Quantification of the protein levels. Data from at least three independent experiments with duplicate determinations are expressed as the mean ± SEM. * P

Techniques Used: Expressing, Western Blot

ERK signaling is key to the effect of α-mangostin on the expression of MMP-2, MMP-9 and E-cadherin. (A) α-mangostin reversed LPS-induced ERK1/2 activation. (B) Quantification of protein levels. (C) The efficacy of ERK siRNA for knocking down ERK protein was demonstrated by western blotting. (D) Cells were treated with LPS (5 μ g/ml), α-mangostin (5 μ M) and/or ERK siRNA for 24 h, then subjected to analyses for cell invasion. (E) BxPC-3 cells were treated with LPS (5 μ g/ml) and/or α-mangostin (5 μ M), with or without ERK siRNA. After 24 h, the E-cadherin, MMP-9 and MMP-2 protein expression levels were detected using western blotting. (F) Quantification of protein levels. Data from at least three independent experiments with duplicate determinations are expressed as the mean ± SEM. * P
Figure Legend Snippet: ERK signaling is key to the effect of α-mangostin on the expression of MMP-2, MMP-9 and E-cadherin. (A) α-mangostin reversed LPS-induced ERK1/2 activation. (B) Quantification of protein levels. (C) The efficacy of ERK siRNA for knocking down ERK protein was demonstrated by western blotting. (D) Cells were treated with LPS (5 μ g/ml), α-mangostin (5 μ M) and/or ERK siRNA for 24 h, then subjected to analyses for cell invasion. (E) BxPC-3 cells were treated with LPS (5 μ g/ml) and/or α-mangostin (5 μ M), with or without ERK siRNA. After 24 h, the E-cadherin, MMP-9 and MMP-2 protein expression levels were detected using western blotting. (F) Quantification of protein levels. Data from at least three independent experiments with duplicate determinations are expressed as the mean ± SEM. * P

Techniques Used: Expressing, Activation Assay, Western Blot

44) Product Images from "PinX1 serves as a potential prognostic indicator for clear cell renal cell carcinoma and inhibits its invasion and metastasis by suppressing MMP-2 via NF-κB-dependent transcription"

Article Title: PinX1 serves as a potential prognostic indicator for clear cell renal cell carcinoma and inhibits its invasion and metastasis by suppressing MMP-2 via NF-κB-dependent transcription

Journal: Oncotarget

doi:

PinX1 inhibits migration and invasion of ccRCC cells by suppressing MMP-2 expression and activity a. Western blotting of PinX1, MMP-2, MMP-9, TIMP-1 and TIMP-2 from ccRCC cells transfected with the PinX1 siRNA or control siRNA. MMP-2 expression was up-regulated independent of TIMP-2 in PinX1 knockdown ccRCC cells. b. Western blotting of PinX1, MMP-2, MMP-9, TIMP-1 and TIMP-2 from ccRCC cells transfected with the pEGFP-C3-PinX1 plasmid or vector control. MMP-2 expression was down-regulated independent of TIMP-2 in PinX1 overexpression ccRCC cells. c. top panel, Gelatin zymography analysis of the enzyme activity of MMP-2 in PinX1 knockdown and control group for both 786-O and ACHN cell lines (gels were incubated for 10 h for 786-O cells and 48 h for ACHN cells). Bottom panel, Gelatin zymography analysis of the enzyme activity of MMP-2 in PinX1 overexpression and control group for both 786-O and ACHN cell lines (gels were incubated for 16 h for 786-O cells and 96 h for ACHN cells). The MMP-2 enzyme activity was significantly enhanced after PinX1 overexpressing in ccRCC cells, conspicuously suppressed after PinX1 knockdown. d. Western blotting of PinX1 and MMP-2 from ccRCC cells transfected with the control siRNA, PinX1 siRNA or co-treated with MMP-2 selective inhibitor I (10 μM). The enhancement of MMP-2 expression regulated by PinX1 knockdown in ccRCC cells was blocked by MMP-2 inhibitor I. e. Gelatin zymography analysis of the enzyme activity of MMP-2 in ccRCC cells transfected with the PinX1 siRNA or co-treated with MMP-2 selective inhibitor I (10 μM). The enhancement of MMP-2 activity regulated by PinX1 knockdown in ccRCC cells was blocked by MMP-2 inhibitor. f. and g. The enhancement of migration and invasion regulated by PinX1 knockdown in ccRCC cells was blocked by MMP-2 inhibitor I. All experiments were carried out in triplicate. Histograms represent means ± SD. ***, P
Figure Legend Snippet: PinX1 inhibits migration and invasion of ccRCC cells by suppressing MMP-2 expression and activity a. Western blotting of PinX1, MMP-2, MMP-9, TIMP-1 and TIMP-2 from ccRCC cells transfected with the PinX1 siRNA or control siRNA. MMP-2 expression was up-regulated independent of TIMP-2 in PinX1 knockdown ccRCC cells. b. Western blotting of PinX1, MMP-2, MMP-9, TIMP-1 and TIMP-2 from ccRCC cells transfected with the pEGFP-C3-PinX1 plasmid or vector control. MMP-2 expression was down-regulated independent of TIMP-2 in PinX1 overexpression ccRCC cells. c. top panel, Gelatin zymography analysis of the enzyme activity of MMP-2 in PinX1 knockdown and control group for both 786-O and ACHN cell lines (gels were incubated for 10 h for 786-O cells and 48 h for ACHN cells). Bottom panel, Gelatin zymography analysis of the enzyme activity of MMP-2 in PinX1 overexpression and control group for both 786-O and ACHN cell lines (gels were incubated for 16 h for 786-O cells and 96 h for ACHN cells). The MMP-2 enzyme activity was significantly enhanced after PinX1 overexpressing in ccRCC cells, conspicuously suppressed after PinX1 knockdown. d. Western blotting of PinX1 and MMP-2 from ccRCC cells transfected with the control siRNA, PinX1 siRNA or co-treated with MMP-2 selective inhibitor I (10 μM). The enhancement of MMP-2 expression regulated by PinX1 knockdown in ccRCC cells was blocked by MMP-2 inhibitor I. e. Gelatin zymography analysis of the enzyme activity of MMP-2 in ccRCC cells transfected with the PinX1 siRNA or co-treated with MMP-2 selective inhibitor I (10 μM). The enhancement of MMP-2 activity regulated by PinX1 knockdown in ccRCC cells was blocked by MMP-2 inhibitor. f. and g. The enhancement of migration and invasion regulated by PinX1 knockdown in ccRCC cells was blocked by MMP-2 inhibitor I. All experiments were carried out in triplicate. Histograms represent means ± SD. ***, P

Techniques Used: Migration, Expressing, Activity Assay, Western Blot, Transfection, Plasmid Preparation, Over Expression, Zymography, Incubation

PinX1 inhibits migration and invasion of ccRCC cells by suppressing MMP-2 expression via NF-κB-dependent transcription a. Western blotting of PinX1 and p65 from ccRCC cells transfected with the PinX1 siRNA or control siRNA. p65 expression was up-regulated in PinX1 knockdown ccRCC cells. b. Western blotting of PinX1 and p65 from ccRCC cells transfected with the pEGFP-C3-PinX1 plasmid or vector control. p65 expression was down-regulated in PinX1 overexpression ccRCC cells. c. RT-PCR determined mRNA levels of PinX1 and p65 in ccRCC cells transfected with PinX1 siRNA or control siRNA. The mRNA level of p65 was increased in PinX1 knockdown ccRCC cells. d. RT-PCR determined mRNA levels of PinX1 and p65 in ccRCC cells transfected with pEGFP-C3-PinX1 plasmid or vector control. The mRNA level of p65 was decreased in PinX1 overexpression ccRCC cells. e and f. Western blotting determined cellular distribution of p65 in 786-O and ACHN cells transfected with control siRNA and PinX1 siRNA. Inhibition of PinX1 significantly increased the nuclear accumulation of p65. g. and h. Western blotting of PinX1, p65 and MMP-2 from ccRCC cells transfected with the control siRNA, PinX1 siRNA, p65 siRNA or co-treated with PinX1 siRNA and p65 siRNA. MMP-2 expression was inhibited by p65 siRNA as well as p65 expression. The enhancement of MMP-2 expression regulated by PinX1 knockdown as well as p65 in ccRCC cells was abolished by p65 siRNA. i. and j. The enhancement of migration and invasion regulated by PinX1 knockdown in ccRCC cells was abolished by p65 siRNA. All experiments were carried out in triplicate. Histograms represent means ± SD. ***, P
Figure Legend Snippet: PinX1 inhibits migration and invasion of ccRCC cells by suppressing MMP-2 expression via NF-κB-dependent transcription a. Western blotting of PinX1 and p65 from ccRCC cells transfected with the PinX1 siRNA or control siRNA. p65 expression was up-regulated in PinX1 knockdown ccRCC cells. b. Western blotting of PinX1 and p65 from ccRCC cells transfected with the pEGFP-C3-PinX1 plasmid or vector control. p65 expression was down-regulated in PinX1 overexpression ccRCC cells. c. RT-PCR determined mRNA levels of PinX1 and p65 in ccRCC cells transfected with PinX1 siRNA or control siRNA. The mRNA level of p65 was increased in PinX1 knockdown ccRCC cells. d. RT-PCR determined mRNA levels of PinX1 and p65 in ccRCC cells transfected with pEGFP-C3-PinX1 plasmid or vector control. The mRNA level of p65 was decreased in PinX1 overexpression ccRCC cells. e and f. Western blotting determined cellular distribution of p65 in 786-O and ACHN cells transfected with control siRNA and PinX1 siRNA. Inhibition of PinX1 significantly increased the nuclear accumulation of p65. g. and h. Western blotting of PinX1, p65 and MMP-2 from ccRCC cells transfected with the control siRNA, PinX1 siRNA, p65 siRNA or co-treated with PinX1 siRNA and p65 siRNA. MMP-2 expression was inhibited by p65 siRNA as well as p65 expression. The enhancement of MMP-2 expression regulated by PinX1 knockdown as well as p65 in ccRCC cells was abolished by p65 siRNA. i. and j. The enhancement of migration and invasion regulated by PinX1 knockdown in ccRCC cells was abolished by p65 siRNA. All experiments were carried out in triplicate. Histograms represent means ± SD. ***, P

Techniques Used: Migration, Expressing, Western Blot, Transfection, Plasmid Preparation, Over Expression, Reverse Transcription Polymerase Chain Reaction, Inhibition

A hypothetic model of PinX1 suppresses MMP-2 expression via NF-κB signaling pathway We presume that PinX1 can inhibit the expression of MMP-2 owing to the suppression of transcriptional activity of NF-κB proteins by direct protein-protein interaction with its G-patch domain.
Figure Legend Snippet: A hypothetic model of PinX1 suppresses MMP-2 expression via NF-κB signaling pathway We presume that PinX1 can inhibit the expression of MMP-2 owing to the suppression of transcriptional activity of NF-κB proteins by direct protein-protein interaction with its G-patch domain.

Techniques Used: Expressing, Activity Assay

45) Product Images from "Anti-proliferative effect of olmesartan on Tenon's capsule fibroblasts"

Article Title: Anti-proliferative effect of olmesartan on Tenon's capsule fibroblasts

Journal: International Journal of Ophthalmology

doi: 10.18240/ijo.2016.05.05

Olmesartan treated-eye presents with lower MMP-2 expression by means of mean density in each group comparing with the controlled eye.
Figure Legend Snippet: Olmesartan treated-eye presents with lower MMP-2 expression by means of mean density in each group comparing with the controlled eye.

Techniques Used: Expressing

The immunohistochemical pictures show the expression of MMP-2 in conjunctiva and subconjunctival tissue of 10×40 magnification
Figure Legend Snippet: The immunohistochemical pictures show the expression of MMP-2 in conjunctiva and subconjunctival tissue of 10×40 magnification

Techniques Used: Immunohistochemistry, Expressing

46) Product Images from "Anti-proliferative effect of olmesartan on Tenon's capsule fibroblasts"

Article Title: Anti-proliferative effect of olmesartan on Tenon's capsule fibroblasts

Journal: International Journal of Ophthalmology

doi: 10.18240/ijo.2016.05.05

Olmesartan treated-eye presents with lower MMP-2 expression by means of mean density in each group comparing with the controlled eye.
Figure Legend Snippet: Olmesartan treated-eye presents with lower MMP-2 expression by means of mean density in each group comparing with the controlled eye.

Techniques Used: Expressing

The immunohistochemical pictures show the expression of MMP-2 in conjunctiva and subconjunctival tissue of 10×40 magnification
Figure Legend Snippet: The immunohistochemical pictures show the expression of MMP-2 in conjunctiva and subconjunctival tissue of 10×40 magnification

Techniques Used: Immunohistochemistry, Expressing

47) Product Images from "Anti-proliferative effect of olmesartan on Tenon's capsule fibroblasts"

Article Title: Anti-proliferative effect of olmesartan on Tenon's capsule fibroblasts

Journal: International Journal of Ophthalmology

doi: 10.18240/ijo.2016.05.05

Olmesartan treated-eye presents with lower MMP-2 expression by means of mean density in each group comparing with the controlled eye.
Figure Legend Snippet: Olmesartan treated-eye presents with lower MMP-2 expression by means of mean density in each group comparing with the controlled eye.

Techniques Used: Expressing

The immunohistochemical pictures show the expression of MMP-2 in conjunctiva and subconjunctival tissue of 10×40 magnification
Figure Legend Snippet: The immunohistochemical pictures show the expression of MMP-2 in conjunctiva and subconjunctival tissue of 10×40 magnification

Techniques Used: Immunohistochemistry, Expressing

48) Product Images from "Anti-proliferative effect of olmesartan on Tenon's capsule fibroblasts"

Article Title: Anti-proliferative effect of olmesartan on Tenon's capsule fibroblasts

Journal: International Journal of Ophthalmology

doi: 10.18240/ijo.2016.05.05

Olmesartan treated-eye presents with lower MMP-2 expression by means of mean density in each group comparing with the controlled eye.
Figure Legend Snippet: Olmesartan treated-eye presents with lower MMP-2 expression by means of mean density in each group comparing with the controlled eye.

Techniques Used: Expressing

The immunohistochemical pictures show the expression of MMP-2 in conjunctiva and subconjunctival tissue of 10×40 magnification
Figure Legend Snippet: The immunohistochemical pictures show the expression of MMP-2 in conjunctiva and subconjunctival tissue of 10×40 magnification

Techniques Used: Immunohistochemistry, Expressing

49) Product Images from "Propofol suppresses proliferation and invasion of glioma cells by upregulating microRNA-218 expression"

Article Title: Propofol suppresses proliferation and invasion of glioma cells by upregulating microRNA-218 expression

Journal: Molecular Medicine Reports

doi: 10.3892/mmr.2015.4014

Western blot analysis showed that propofol treatment downregulated matrix metalloproteinase-2 (MMP-2) expression. (A) The protein levels of MMP-2 in U373 cells were markedly decreased following propofol treatment. (B) Statistical analysis of MMP-2 protein levels. *P
Figure Legend Snippet: Western blot analysis showed that propofol treatment downregulated matrix metalloproteinase-2 (MMP-2) expression. (A) The protein levels of MMP-2 in U373 cells were markedly decreased following propofol treatment. (B) Statistical analysis of MMP-2 protein levels. *P

Techniques Used: Western Blot, Expressing

Overexpression of microRNA (miR)-218 can suppress matrix metalloproteinase (MMP)-2 expression. (A) Transfection of U373 human glioma cells with miR-218 precursor significantly elevated the expression levels of miR-218. (B) Western blot analysis indicated that transfection with miR-218 precursor decreased MMP-2 protein expression levels. *P
Figure Legend Snippet: Overexpression of microRNA (miR)-218 can suppress matrix metalloproteinase (MMP)-2 expression. (A) Transfection of U373 human glioma cells with miR-218 precursor significantly elevated the expression levels of miR-218. (B) Western blot analysis indicated that transfection with miR-218 precursor decreased MMP-2 protein expression levels. *P

Techniques Used: Over Expression, Expressing, Transfection, Western Blot

50) Product Images from "Plaque Rupture is a Determinant of Vascular Events in Carotid Artery Atherosclerotic Disease: Involvement of Matrix Metalloproteinases 2 and 9"

Article Title: Plaque Rupture is a Determinant of Vascular Events in Carotid Artery Atherosclerotic Disease: Involvement of Matrix Metalloproteinases 2 and 9

Journal: Journal of Clinical Neurology (Seoul, Korea)

doi: 10.3988/jcn.2011.7.2.69

Immunofluorescence staining illustrating the expressions of MMP-2 and MMP-9 produced by macrophages in carotid plaques. A and D: MMP-2 and MMP-9 were visualized with secondary antibodies conjugated to Texas Red (red). B and E: The nuclei were stained with DAPI (blue). C and F: MMPs colocalized with the cytoplasm in the macrophages. The white boxes in each image are magnified. Scale bar=100 µm. MMP: matrix metalloproteinase.
Figure Legend Snippet: Immunofluorescence staining illustrating the expressions of MMP-2 and MMP-9 produced by macrophages in carotid plaques. A and D: MMP-2 and MMP-9 were visualized with secondary antibodies conjugated to Texas Red (red). B and E: The nuclei were stained with DAPI (blue). C and F: MMPs colocalized with the cytoplasm in the macrophages. The white boxes in each image are magnified. Scale bar=100 µm. MMP: matrix metalloproteinase.

Techniques Used: Immunofluorescence, Staining, Produced

Carotid specimens that were immunohistochemically positive for MMP-2 (A) and MMP-9 (B) were also strongly positive for CD68 (C) but negative for SMA (D). MMP: matrix metalloproteinase, SMA: smooth-muscle actin.
Figure Legend Snippet: Carotid specimens that were immunohistochemically positive for MMP-2 (A) and MMP-9 (B) were also strongly positive for CD68 (C) but negative for SMA (D). MMP: matrix metalloproteinase, SMA: smooth-muscle actin.

Techniques Used:

51) Product Images from "CIP4 Promotes Lung Adenocarcinoma Metastasis and Is Associated with Poor Prognosis"

Article Title: CIP4 Promotes Lung Adenocarcinoma Metastasis and Is Associated with Poor Prognosis

Journal: Oncogene

doi: 10.1038/onc.2014.280

CIP4 promotes NSCLC tumor metastasis in mice. (a) Representative images of subcutaneous H1299 V and KD1 tumors 5 weeks after injection of Rag2 −/− :IL-2Rγc −/− mice (scale bar is 2.5 mm). (b) Graph depicts primary tumor mass (mean ± SEM) for mice injected with H1299 V and KD1 cells (N=13 mice/group from 3 experiments). (c) CIP4 and MMP-2 expression in H1299 V and KD1 tumor homogenates was determined by IB analysis. Protein positions are indicated on the right. (d) Relative expression of MMP-2 and Zeb1 transcripts in H1299 V and KD1 tumors. Total RNA was isolated and analyzed by qRT-PCR using MMP-2 or Zeb1 specific primers and normalized to GAPDH expression. (e) Spontaneous metastases to lungs in mice harboring H1299 V and KD1 subcutaneous tumors were detected by GFP fluorescence in whole lung tissue (top), and scored from H E stained lung sections (bottom; metastases are indicated by arrows; scale bar indicates 50 μm). (f) Graph depicts numbers of lung metastases (mean ± SEM) that were counted in a blinded fashion (N=13 mice/group from 3 experiments). (g) Experimental metastasis assays with H1299 V and KD1 cells injected in the spleen were performed and representative images of H E stained sections are shown for liver (top) and lung (lower; scale bar indicates 50 μm). (h) Graph depicts number of liver and lung metastases after splenic injection of H1299 V and KD1 cells (mean ± SEM; N=9 mice/group from 3 experiments; * indicates significant differences between cell lines ( P
Figure Legend Snippet: CIP4 promotes NSCLC tumor metastasis in mice. (a) Representative images of subcutaneous H1299 V and KD1 tumors 5 weeks after injection of Rag2 −/− :IL-2Rγc −/− mice (scale bar is 2.5 mm). (b) Graph depicts primary tumor mass (mean ± SEM) for mice injected with H1299 V and KD1 cells (N=13 mice/group from 3 experiments). (c) CIP4 and MMP-2 expression in H1299 V and KD1 tumor homogenates was determined by IB analysis. Protein positions are indicated on the right. (d) Relative expression of MMP-2 and Zeb1 transcripts in H1299 V and KD1 tumors. Total RNA was isolated and analyzed by qRT-PCR using MMP-2 or Zeb1 specific primers and normalized to GAPDH expression. (e) Spontaneous metastases to lungs in mice harboring H1299 V and KD1 subcutaneous tumors were detected by GFP fluorescence in whole lung tissue (top), and scored from H E stained lung sections (bottom; metastases are indicated by arrows; scale bar indicates 50 μm). (f) Graph depicts numbers of lung metastases (mean ± SEM) that were counted in a blinded fashion (N=13 mice/group from 3 experiments). (g) Experimental metastasis assays with H1299 V and KD1 cells injected in the spleen were performed and representative images of H E stained sections are shown for liver (top) and lung (lower; scale bar indicates 50 μm). (h) Graph depicts number of liver and lung metastases after splenic injection of H1299 V and KD1 cells (mean ± SEM; N=9 mice/group from 3 experiments; * indicates significant differences between cell lines ( P

Techniques Used: Mouse Assay, Injection, Expressing, Isolation, Quantitative RT-PCR, Fluorescence, Staining

CIP4 promotes MMP-2 expression in NSCLC cells. (a) Concentrations of MMP-2 and MMP-9 proteins in CM from H1299 V and KD1 cells. Values were calculated by Eve Technologies using a human MMP multiplex assay. (b) Gelatin zymography of CM from H1299 V and KD1 cells. 10 μg protein per lane were analyzed. Positions of MMP-2 and MMP-9 activity are indicated on the right with size markers on the left. (c) Quantification of MMP-2 and MMP-9 activity by gelatin zymography. Bands of gelatin digestion for MMP-2 and MMP-9 from H1299 V and KD1 CMs were measured by densitometry. (d) IB analysis of MMP-2 in H1299 V and KD1 cell lysates (10 μg protein per lane). Positions of MMP-2, CIP4 and β-actin are shown on the right with size markers on the left. (e) Relative expression of MMP-2 transcripts in H1299 V and KD1 cells. Total RNA was isolated and analyzed by qRT-PCR using MMP-2 specific primers and normalized to GAPDH expression. All experiments were performed 3 times in triplicate (Graphs are shown as the mean ± SEM, * indicates significant differences between cell lines ( P
Figure Legend Snippet: CIP4 promotes MMP-2 expression in NSCLC cells. (a) Concentrations of MMP-2 and MMP-9 proteins in CM from H1299 V and KD1 cells. Values were calculated by Eve Technologies using a human MMP multiplex assay. (b) Gelatin zymography of CM from H1299 V and KD1 cells. 10 μg protein per lane were analyzed. Positions of MMP-2 and MMP-9 activity are indicated on the right with size markers on the left. (c) Quantification of MMP-2 and MMP-9 activity by gelatin zymography. Bands of gelatin digestion for MMP-2 and MMP-9 from H1299 V and KD1 CMs were measured by densitometry. (d) IB analysis of MMP-2 in H1299 V and KD1 cell lysates (10 μg protein per lane). Positions of MMP-2, CIP4 and β-actin are shown on the right with size markers on the left. (e) Relative expression of MMP-2 transcripts in H1299 V and KD1 cells. Total RNA was isolated and analyzed by qRT-PCR using MMP-2 specific primers and normalized to GAPDH expression. All experiments were performed 3 times in triplicate (Graphs are shown as the mean ± SEM, * indicates significant differences between cell lines ( P

Techniques Used: Expressing, Multiplex Assay, Zymography, Activity Assay, Isolation, Quantitative RT-PCR

52) Product Images from "Combining two potential causes of metalloproteinase secretion causes abdominal aortic aneurysms in rats: a new experimental model"

Article Title: Combining two potential causes of metalloproteinase secretion causes abdominal aortic aneurysms in rats: a new experimental model

Journal: International Journal of Experimental Pathology

doi: 10.1111/j.1365-2613.2010.00746.x

Immunohistochemical localization of MMP-9 and MMP-2 in aneurismal wall on day 3 after surgery are show. (a) MMP-9 showed strong expression throughout the remodelling wall, especially within neutrophils (arrowheads), macrophages (arrow) and myofibroblasts
Figure Legend Snippet: Immunohistochemical localization of MMP-9 and MMP-2 in aneurismal wall on day 3 after surgery are show. (a) MMP-9 showed strong expression throughout the remodelling wall, especially within neutrophils (arrowheads), macrophages (arrow) and myofibroblasts

Techniques Used: Immunohistochemistry, Expressing

Bar graphs show the densitometric analyses of the MMP-2 and MMP-9 activities during the experiment. (a) Note the elevated levels of the MMP-9 in the Aneurysm group, starting at the first dps and remaining elevated at similar levels until the end of the
Figure Legend Snippet: Bar graphs show the densitometric analyses of the MMP-2 and MMP-9 activities during the experiment. (a) Note the elevated levels of the MMP-9 in the Aneurysm group, starting at the first dps and remaining elevated at similar levels until the end of the

Techniques Used:

Representative SDS–PAGE gelatin zymography of aortic samples (a–d), showing increased expression of MMP-9, identified as a 92 kDa band, and latent (72 kDa) and active forms (64 kDa) of MMP-2 in the aneurysms extracts (AAA) in all days
Figure Legend Snippet: Representative SDS–PAGE gelatin zymography of aortic samples (a–d), showing increased expression of MMP-9, identified as a 92 kDa band, and latent (72 kDa) and active forms (64 kDa) of MMP-2 in the aneurysms extracts (AAA) in all days

Techniques Used: SDS Page, Zymography, Expressing

53) Product Images from "Intervention of human breast cell carcinogenesis chronically induced by 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine"

Article Title: Intervention of human breast cell carcinogenesis chronically induced by 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine

Journal: Carcinogenesis

doi: 10.1093/carcin/bgs097

H-Ras, the ERK pathway, Nox-1 and ROS in modulation of cancer-associated properties. ( A and B ) P-20 and MCF10A-Ras cells were transfected with control siRNA (CR) and validated Nox-1- or H-Ras-specific siRNAs for 40 h. ( C ) 10A and P-20 cells were treated with 10 μmol/l U0126 (U0) for 48 h. ( D ) P-20 cells were treated with 10 μmol/l U0, 0.1 μmol/l DPI or 5 mmol/l NAC for 48 h. (A to C) Cell lysates were prepared and analyzed by western immunoblotting to detect levels of H-Ras, p-Erk1/2, Erk1/2, Nox-1, TNF-α, MMP-2, MMP-9 and E-cadherin, with β-actin as a control, and these levels were quantified by densitometry. The levels of H-Ras, Nox-1, TNF-α, MMP-2, MMP-9 and E-cadherin were calculated by normalizing with the level of β-actin, the level set in control cells as 1 (X, arbitrary unit). The level of specific phosphorylation of Erk1/2 (p/Erk) was calculated by normalizing the level of p-Erk1/2 with the level of Erk1/2, then the level set in control cells as 1 (X, arbitrary unit). (A, B and D) Relative proliferation was determined and normalized by the value of 5-bromo-2′-deoxyuridine detected in control siRNA-transfected counterpart cells (A and B) or untreated 10A control cells (D), set as 100%. Relative ROS levels were measured with chloromethyl dichlorodihydrofluorescein diacetate labeling and normalized by fluorescence intensity determined in control cells, set as 100%. (D) Migratory and invasive activities were determined by counting the numbers of cells translocated through a polycarbonate filter without or with coated Matrigel, respectively, in 10 arbitrary visual fields; relative migratory and invasive activities were normalized by the value of migrated or invaded cells detected in untreated cells, respectively, set as 100%. To determine serum-independent non-adherent growth (SINAG), cells were seeded in non-adherent cultures in the absence and presence of 10 μmol/l U0, 0.1 μmol/l DPI or 5 mmol/l NAC for 10 days; then, mammospheres (≥0.1 mm diameter) were counted. Relative mammosphere formations were normalized by the number of mammospheres formed in untreated cells, set as 100%. Columns, mean of triplicates; bars, standard deviation. The Student’s t test was used to analyze statistical significance, indicated by ** P
Figure Legend Snippet: H-Ras, the ERK pathway, Nox-1 and ROS in modulation of cancer-associated properties. ( A and B ) P-20 and MCF10A-Ras cells were transfected with control siRNA (CR) and validated Nox-1- or H-Ras-specific siRNAs for 40 h. ( C ) 10A and P-20 cells were treated with 10 μmol/l U0126 (U0) for 48 h. ( D ) P-20 cells were treated with 10 μmol/l U0, 0.1 μmol/l DPI or 5 mmol/l NAC for 48 h. (A to C) Cell lysates were prepared and analyzed by western immunoblotting to detect levels of H-Ras, p-Erk1/2, Erk1/2, Nox-1, TNF-α, MMP-2, MMP-9 and E-cadherin, with β-actin as a control, and these levels were quantified by densitometry. The levels of H-Ras, Nox-1, TNF-α, MMP-2, MMP-9 and E-cadherin were calculated by normalizing with the level of β-actin, the level set in control cells as 1 (X, arbitrary unit). The level of specific phosphorylation of Erk1/2 (p/Erk) was calculated by normalizing the level of p-Erk1/2 with the level of Erk1/2, then the level set in control cells as 1 (X, arbitrary unit). (A, B and D) Relative proliferation was determined and normalized by the value of 5-bromo-2′-deoxyuridine detected in control siRNA-transfected counterpart cells (A and B) or untreated 10A control cells (D), set as 100%. Relative ROS levels were measured with chloromethyl dichlorodihydrofluorescein diacetate labeling and normalized by fluorescence intensity determined in control cells, set as 100%. (D) Migratory and invasive activities were determined by counting the numbers of cells translocated through a polycarbonate filter without or with coated Matrigel, respectively, in 10 arbitrary visual fields; relative migratory and invasive activities were normalized by the value of migrated or invaded cells detected in untreated cells, respectively, set as 100%. To determine serum-independent non-adherent growth (SINAG), cells were seeded in non-adherent cultures in the absence and presence of 10 μmol/l U0, 0.1 μmol/l DPI or 5 mmol/l NAC for 10 days; then, mammospheres (≥0.1 mm diameter) were counted. Relative mammosphere formations were normalized by the number of mammospheres formed in untreated cells, set as 100%. Columns, mean of triplicates; bars, standard deviation. The Student’s t test was used to analyze statistical significance, indicated by ** P

Techniques Used: Transfection, Western Blot, Labeling, Fluorescence, Standard Deviation

Constitutive targeted endpoints in progressive carcinogenesis induced by PhIP. MCF10A (10A) cells were repeatedly exposed to 10 nmol/l PhIP for 10 and 20 cycles, resulting in the MCF10A-10nM-P10 (P-10) and -P20 (P-20) cell lines, respectively. MCF10A cells were stably transfected to ectopically express oncogenic H-Ras, resulting in the MCF10A-Ras cell line (Ras). ( A ) Cells were maintained in low-mitogen medium for 10 days, seeded in soft agar for 14 days and seeded in Matrigel for 14 days to determine cellular acquisition of reduced dependence on growth factors (RDGF), anchorage-independent growth (AIG) and acinar-conformational disruption (ACD), respectively. Cell colonies (≥0.5 mm diameter) grown in low-mitogen medium and soft agar were counted. Regular and irregular spheroids in Matrigel were counted, and the percentage of irregular spheroids was calculated. Cell proliferation was determined; relative cell growth rate was normalized by the value of 5-bromo-2′-deoxyuridine detected in 10A cells, set as 100%. Migratory and invasive activities were determined by counting the numbers of cells translocated through a polycarbonate filter without or with coated Matrigel, respectively in 10 arbitrary visual fields. ( B ) Cell lysates were analyzed by western immunoblotting using specific antibodies to detect levels of H-Ras, phosphorylated Erk1/2 (p-Erk1/2), Erk1/2, Nox-1, TNF-α, MMP-2, MMP-9 and E-cadherin, with β-actin as a control. The levels of H-Ras, Nox-1, TNF-α, MMP-2, MMP-9 and E-cadherin were calculated by normalizing with the level of β-actin, the level set in 10A cells as 1 (X, arbitrary unit). The level of specific phosphorylation of Erk1/2 (p/Erk) was calculated by normalizing the level of p-Erk1/2 with the level of Erk1/2, then the level set in 10A cells as 1 (X, arbitrary unit). ROS levels were measured with chloromethyl dichlorodihydrofluorescein diacetate labeling; relative level of ROS, as fold induction (X, arbitrary unit), was normalized by the level determined in 10A cells, set as 1. Total RNAs (RNA) were isolated and analyzed by PCR with specific primers to determine relative gene expression levels of H-Ras with β-actin as a control. ( C ) To determine serum-independent non-adherent growth (SINAG), cells were seeded in non-adherent cultures for 10 days and mammospheres (≥0.1 mm diameter) were counted. Mammospheres were collected and trypsinized, and ALDH-expressing (ALDH+) cell population (%) was measured by flow cytometry. Columns, mean of triplicates; bars, standard deviation. The Student’s t test was used to analyze statistical significance, indicated by * P
Figure Legend Snippet: Constitutive targeted endpoints in progressive carcinogenesis induced by PhIP. MCF10A (10A) cells were repeatedly exposed to 10 nmol/l PhIP for 10 and 20 cycles, resulting in the MCF10A-10nM-P10 (P-10) and -P20 (P-20) cell lines, respectively. MCF10A cells were stably transfected to ectopically express oncogenic H-Ras, resulting in the MCF10A-Ras cell line (Ras). ( A ) Cells were maintained in low-mitogen medium for 10 days, seeded in soft agar for 14 days and seeded in Matrigel for 14 days to determine cellular acquisition of reduced dependence on growth factors (RDGF), anchorage-independent growth (AIG) and acinar-conformational disruption (ACD), respectively. Cell colonies (≥0.5 mm diameter) grown in low-mitogen medium and soft agar were counted. Regular and irregular spheroids in Matrigel were counted, and the percentage of irregular spheroids was calculated. Cell proliferation was determined; relative cell growth rate was normalized by the value of 5-bromo-2′-deoxyuridine detected in 10A cells, set as 100%. Migratory and invasive activities were determined by counting the numbers of cells translocated through a polycarbonate filter without or with coated Matrigel, respectively in 10 arbitrary visual fields. ( B ) Cell lysates were analyzed by western immunoblotting using specific antibodies to detect levels of H-Ras, phosphorylated Erk1/2 (p-Erk1/2), Erk1/2, Nox-1, TNF-α, MMP-2, MMP-9 and E-cadherin, with β-actin as a control. The levels of H-Ras, Nox-1, TNF-α, MMP-2, MMP-9 and E-cadherin were calculated by normalizing with the level of β-actin, the level set in 10A cells as 1 (X, arbitrary unit). The level of specific phosphorylation of Erk1/2 (p/Erk) was calculated by normalizing the level of p-Erk1/2 with the level of Erk1/2, then the level set in 10A cells as 1 (X, arbitrary unit). ROS levels were measured with chloromethyl dichlorodihydrofluorescein diacetate labeling; relative level of ROS, as fold induction (X, arbitrary unit), was normalized by the level determined in 10A cells, set as 1. Total RNAs (RNA) were isolated and analyzed by PCR with specific primers to determine relative gene expression levels of H-Ras with β-actin as a control. ( C ) To determine serum-independent non-adherent growth (SINAG), cells were seeded in non-adherent cultures for 10 days and mammospheres (≥0.1 mm diameter) were counted. Mammospheres were collected and trypsinized, and ALDH-expressing (ALDH+) cell population (%) was measured by flow cytometry. Columns, mean of triplicates; bars, standard deviation. The Student’s t test was used to analyze statistical significance, indicated by * P

Techniques Used: Stable Transfection, Transfection, Western Blot, Labeling, Isolation, Polymerase Chain Reaction, Expressing, Flow Cytometry, Cytometry, Standard Deviation

54) Product Images from "CircularRNA_104670 plays a critical role in intervertebral disc degeneration by functioning as a ceRNA"

Article Title: CircularRNA_104670 plays a critical role in intervertebral disc degeneration by functioning as a ceRNA

Journal: Experimental & Molecular Medicine

doi: 10.1038/s12276-018-0125-y

circRNA_104670 promotes the expression of MMP-2 and inhibits the expression of collagen II. a , b Immunofluorescence staining for collagen II after transfection with si-circRNA_104670 or co-transfection with si-circRNA_104670 and si-miRNA-17-3p, scale bar, 40 μm. c , d Immunofluorescence staining for MMP-2 after transfection with si-circRNA_104670 or co-transfection with si-circRNA_104670 and si-miRNA-17-3p, scale bar, 40 μm. e – g Collagen II and MMP-2 protein expression levels were analyzed by Western blotting. GAPDH was used as a loading control. The presented values are the means ± SD. of three different preparations, * p
Figure Legend Snippet: circRNA_104670 promotes the expression of MMP-2 and inhibits the expression of collagen II. a , b Immunofluorescence staining for collagen II after transfection with si-circRNA_104670 or co-transfection with si-circRNA_104670 and si-miRNA-17-3p, scale bar, 40 μm. c , d Immunofluorescence staining for MMP-2 after transfection with si-circRNA_104670 or co-transfection with si-circRNA_104670 and si-miRNA-17-3p, scale bar, 40 μm. e – g Collagen II and MMP-2 protein expression levels were analyzed by Western blotting. GAPDH was used as a loading control. The presented values are the means ± SD. of three different preparations, * p

Techniques Used: Expressing, Immunofluorescence, Staining, Transfection, Cotransfection, Western Blot

CircRNA 104670 serves as a sponge for the miR-17-3p and MMP-2 is directly targeted by miR-17-3p. a The binding region of miRNAs in circRNA_104670 3′UTR are shown. b A luciferase reporter assay confirmed that miR-17-3p, miR-22-5p, miR-96-3p, and miR-433-3p reduce the luciferase intensity by more than 40%. c , d A pmiR-RB-Report vector with or without the 3′UTR sequence of circRNA_104670 was co-transfected with miR-17-3p or a negative control oligonucleotide; RLU of hRluc and hLuc+ were constructed, and the hRluc values were normalized to the corresponding hLuc+ values, ** p
Figure Legend Snippet: CircRNA 104670 serves as a sponge for the miR-17-3p and MMP-2 is directly targeted by miR-17-3p. a The binding region of miRNAs in circRNA_104670 3′UTR are shown. b A luciferase reporter assay confirmed that miR-17-3p, miR-22-5p, miR-96-3p, and miR-433-3p reduce the luciferase intensity by more than 40%. c , d A pmiR-RB-Report vector with or without the 3′UTR sequence of circRNA_104670 was co-transfected with miR-17-3p or a negative control oligonucleotide; RLU of hRluc and hLuc+ were constructed, and the hRluc values were normalized to the corresponding hLuc+ values, ** p

Techniques Used: Binding Assay, Luciferase, Reporter Assay, Plasmid Preparation, Sequencing, Transfection, Negative Control, Construct

55) Product Images from "Plaque Rupture is a Determinant of Vascular Events in Carotid Artery Atherosclerotic Disease: Involvement of Matrix Metalloproteinases 2 and 9"

Article Title: Plaque Rupture is a Determinant of Vascular Events in Carotid Artery Atherosclerotic Disease: Involvement of Matrix Metalloproteinases 2 and 9

Journal: Journal of Clinical Neurology (Seoul, Korea)

doi: 10.3988/jcn.2011.7.2.69

Immunofluorescence staining illustrating the expressions of MMP-2 and MMP-9 produced by macrophages in carotid plaques. A and D: MMP-2 and MMP-9 were visualized with secondary antibodies conjugated to Texas Red (red). B and E: The nuclei were stained with DAPI (blue). C and F: MMPs colocalized with the cytoplasm in the macrophages. The white boxes in each image are magnified. Scale bar=100 µm. MMP: matrix metalloproteinase.
Figure Legend Snippet: Immunofluorescence staining illustrating the expressions of MMP-2 and MMP-9 produced by macrophages in carotid plaques. A and D: MMP-2 and MMP-9 were visualized with secondary antibodies conjugated to Texas Red (red). B and E: The nuclei were stained with DAPI (blue). C and F: MMPs colocalized with the cytoplasm in the macrophages. The white boxes in each image are magnified. Scale bar=100 µm. MMP: matrix metalloproteinase.

Techniques Used: Immunofluorescence, Staining, Produced

Carotid specimens that were immunohistochemically positive for MMP-2 (A) and MMP-9 (B) were also strongly positive for CD68 (C) but negative for SMA (D). MMP: matrix metalloproteinase, SMA: smooth-muscle actin.
Figure Legend Snippet: Carotid specimens that were immunohistochemically positive for MMP-2 (A) and MMP-9 (B) were also strongly positive for CD68 (C) but negative for SMA (D). MMP: matrix metalloproteinase, SMA: smooth-muscle actin.

Techniques Used:

56) Product Images from "Anthocyanins from Vitis coignetiae Pulliat Inhibit Cancer Invasion and Epithelial-Mesenchymal Transition, but These Effects Can Be Attenuated by Tumor Necrosis Factor in Human Uterine Cervical Cancer HeLa Cells"

Article Title: Anthocyanins from Vitis coignetiae Pulliat Inhibit Cancer Invasion and Epithelial-Mesenchymal Transition, but These Effects Can Be Attenuated by Tumor Necrosis Factor in Human Uterine Cervical Cancer HeLa Cells

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

doi: 10.1155/2013/503043

Effects of AIMs on cancer cell migration and invasion in HeLa cells treated with TNF. (a) Cells were serum-starved for 24 hrs with or without AIMs (400 μ g/mL). Cells (5 × 10 4 cells) were loaded onto precoated Matrigel 24-well invasion chambers in the presence or absence of TNF (10 ng/mL). The Matrigel invasion chambers were incubated for 24 h. (×400; the length of scale bar, 40 μ m). (b) Cells were grown to 100% confluency on 30 mm cell culture dishes coated with collagen and then serum-starved for 24 hrs with or without TNF (10 ng/mL) and/or AIMs (400 μ g/mL) for 24 h. A linear scratch was made on the culture dish through the cell layer using a pipette tip. After washing with PBS, serum-free media with or without AIMs were added. Photographs of the etched area in a Petri dish were taken at the interval of 0 h, 12 h, 24 h, and 48 h after the scratch to evaluate cell movement into the wounded area. Data were representative of three independent experiments. (c) MMP-2 and MMP-9 protein levels were measured by gelatin zymography. Cells were incubated for 48 h without or with AIMs (25–400 μ g/mL). Values represent means ± SD from three indepe ndent experiments. * P
Figure Legend Snippet: Effects of AIMs on cancer cell migration and invasion in HeLa cells treated with TNF. (a) Cells were serum-starved for 24 hrs with or without AIMs (400 μ g/mL). Cells (5 × 10 4 cells) were loaded onto precoated Matrigel 24-well invasion chambers in the presence or absence of TNF (10 ng/mL). The Matrigel invasion chambers were incubated for 24 h. (×400; the length of scale bar, 40 μ m). (b) Cells were grown to 100% confluency on 30 mm cell culture dishes coated with collagen and then serum-starved for 24 hrs with or without TNF (10 ng/mL) and/or AIMs (400 μ g/mL) for 24 h. A linear scratch was made on the culture dish through the cell layer using a pipette tip. After washing with PBS, serum-free media with or without AIMs were added. Photographs of the etched area in a Petri dish were taken at the interval of 0 h, 12 h, 24 h, and 48 h after the scratch to evaluate cell movement into the wounded area. Data were representative of three independent experiments. (c) MMP-2 and MMP-9 protein levels were measured by gelatin zymography. Cells were incubated for 48 h without or with AIMs (25–400 μ g/mL). Values represent means ± SD from three indepe ndent experiments. * P

Techniques Used: Migration, Incubation, Cell Culture, Transferring, Zymography

Dose-dependent inhibitory effects of AIMs on HeLa cell proliferation and invasion. (a) Growth inhibition of HeLa cells. Cells were seeded at 5 × 10 4 cells/mL and treated with AIMs for 24 h with the indicated concentrations. (b) Effects on invasion of HeLa cells. The cells (5 × 10 4 cells) were loaded on precoated Matrigel 24-well invasion chambers (BD Biosciences) in the presence or absence of AIMs (25–400 μ g/mL). HeLa cells were treated with AIMs for 18 hr in a Matrigel-coated transwell. (c) MMP-2 and MMP-9 protein levels were measured by gelatin zymography. Cells were incubated for 48 h without or with AIMs (25–400 μ g/mL). Values represent means ± SD from three independent experiments. * P
Figure Legend Snippet: Dose-dependent inhibitory effects of AIMs on HeLa cell proliferation and invasion. (a) Growth inhibition of HeLa cells. Cells were seeded at 5 × 10 4 cells/mL and treated with AIMs for 24 h with the indicated concentrations. (b) Effects on invasion of HeLa cells. The cells (5 × 10 4 cells) were loaded on precoated Matrigel 24-well invasion chambers (BD Biosciences) in the presence or absence of AIMs (25–400 μ g/mL). HeLa cells were treated with AIMs for 18 hr in a Matrigel-coated transwell. (c) MMP-2 and MMP-9 protein levels were measured by gelatin zymography. Cells were incubated for 48 h without or with AIMs (25–400 μ g/mL). Values represent means ± SD from three independent experiments. * P

Techniques Used: Inhibition, Zymography, Incubation

57) Product Images from "Anti-proliferative effect of olmesartan on Tenon's capsule fibroblasts"

Article Title: Anti-proliferative effect of olmesartan on Tenon's capsule fibroblasts

Journal: International Journal of Ophthalmology

doi: 10.18240/ijo.2016.05.05

Olmesartan treated-eye presents with lower MMP-2 expression by means of mean density in each group comparing with the controlled eye.
Figure Legend Snippet: Olmesartan treated-eye presents with lower MMP-2 expression by means of mean density in each group comparing with the controlled eye.

Techniques Used: Expressing

The immunohistochemical pictures show the expression of MMP-2 in conjunctiva and subconjunctival tissue of 10×40 magnification
Figure Legend Snippet: The immunohistochemical pictures show the expression of MMP-2 in conjunctiva and subconjunctival tissue of 10×40 magnification

Techniques Used: Immunohistochemistry, Expressing

58) Product Images from "Doxorubicin conjugated with a trastuzumab epitope and an MMP-2 sensitive peptide linker for the treatment of HER2-positive breast cancer"

Article Title: Doxorubicin conjugated with a trastuzumab epitope and an MMP-2 sensitive peptide linker for the treatment of HER2-positive breast cancer

Journal: Drug Delivery

doi: 10.1080/10717544.2018.1435746

Molecular mechanism of MAHNP-DOX. MAHNP-DOX is cleaved extracellularly by MMP-2 and then DOX enters the cells, while the anti-HER2 peptide AHNP binds to HER2, inhibiting the HER2/PI3K/AKT signaling pathway and leading to cell arrest during G1 phase.
Figure Legend Snippet: Molecular mechanism of MAHNP-DOX. MAHNP-DOX is cleaved extracellularly by MMP-2 and then DOX enters the cells, while the anti-HER2 peptide AHNP binds to HER2, inhibiting the HER2/PI3K/AKT signaling pathway and leading to cell arrest during G1 phase.

Techniques Used:

Cytotoxicity profiles of free DOX, MAHNP-DOX and MAHNP-DOX with MMP-2 inhibitor pretreatment of BT474 and SKBR3 cells ( n = 3).
Figure Legend Snippet: Cytotoxicity profiles of free DOX, MAHNP-DOX and MAHNP-DOX with MMP-2 inhibitor pretreatment of BT474 and SKBR3 cells ( n = 3).

Techniques Used:

59) Product Images from "Plaque Rupture is a Determinant of Vascular Events in Carotid Artery Atherosclerotic Disease: Involvement of Matrix Metalloproteinases 2 and 9"

Article Title: Plaque Rupture is a Determinant of Vascular Events in Carotid Artery Atherosclerotic Disease: Involvement of Matrix Metalloproteinases 2 and 9

Journal: Journal of Clinical Neurology (Seoul, Korea)

doi: 10.3988/jcn.2011.7.2.69

Immunofluorescence staining illustrating the expressions of MMP-2 and MMP-9 produced by macrophages in carotid plaques. A and D: MMP-2 and MMP-9 were visualized with secondary antibodies conjugated to Texas Red (red). B and E: The nuclei were stained with DAPI (blue). C and F: MMPs colocalized with the cytoplasm in the macrophages. The white boxes in each image are magnified. Scale bar=100 µm. MMP: matrix metalloproteinase.
Figure Legend Snippet: Immunofluorescence staining illustrating the expressions of MMP-2 and MMP-9 produced by macrophages in carotid plaques. A and D: MMP-2 and MMP-9 were visualized with secondary antibodies conjugated to Texas Red (red). B and E: The nuclei were stained with DAPI (blue). C and F: MMPs colocalized with the cytoplasm in the macrophages. The white boxes in each image are magnified. Scale bar=100 µm. MMP: matrix metalloproteinase.

Techniques Used: Immunofluorescence, Staining, Produced

Carotid specimens that were immunohistochemically positive for MMP-2 (A) and MMP-9 (B) were also strongly positive for CD68 (C) but negative for SMA (D). MMP: matrix metalloproteinase, SMA: smooth-muscle actin.
Figure Legend Snippet: Carotid specimens that were immunohistochemically positive for MMP-2 (A) and MMP-9 (B) were also strongly positive for CD68 (C) but negative for SMA (D). MMP: matrix metalloproteinase, SMA: smooth-muscle actin.

Techniques Used:

60) Product Images from "Plaque Rupture is a Determinant of Vascular Events in Carotid Artery Atherosclerotic Disease: Involvement of Matrix Metalloproteinases 2 and 9"

Article Title: Plaque Rupture is a Determinant of Vascular Events in Carotid Artery Atherosclerotic Disease: Involvement of Matrix Metalloproteinases 2 and 9

Journal: Journal of Clinical Neurology (Seoul, Korea)

doi: 10.3988/jcn.2011.7.2.69

Immunofluorescence staining illustrating the expressions of MMP-2 and MMP-9 produced by macrophages in carotid plaques. A and D: MMP-2 and MMP-9 were visualized with secondary antibodies conjugated to Texas Red (red). B and E: The nuclei were stained with DAPI (blue). C and F: MMPs colocalized with the cytoplasm in the macrophages. The white boxes in each image are magnified. Scale bar=100 µm. MMP: matrix metalloproteinase.
Figure Legend Snippet: Immunofluorescence staining illustrating the expressions of MMP-2 and MMP-9 produced by macrophages in carotid plaques. A and D: MMP-2 and MMP-9 were visualized with secondary antibodies conjugated to Texas Red (red). B and E: The nuclei were stained with DAPI (blue). C and F: MMPs colocalized with the cytoplasm in the macrophages. The white boxes in each image are magnified. Scale bar=100 µm. MMP: matrix metalloproteinase.

Techniques Used: Immunofluorescence, Staining, Produced

Carotid specimens that were immunohistochemically positive for MMP-2 (A) and MMP-9 (B) were also strongly positive for CD68 (C) but negative for SMA (D). MMP: matrix metalloproteinase, SMA: smooth-muscle actin.
Figure Legend Snippet: Carotid specimens that were immunohistochemically positive for MMP-2 (A) and MMP-9 (B) were also strongly positive for CD68 (C) but negative for SMA (D). MMP: matrix metalloproteinase, SMA: smooth-muscle actin.

Techniques Used:

61) Product Images from "Plaque Rupture is a Determinant of Vascular Events in Carotid Artery Atherosclerotic Disease: Involvement of Matrix Metalloproteinases 2 and 9"

Article Title: Plaque Rupture is a Determinant of Vascular Events in Carotid Artery Atherosclerotic Disease: Involvement of Matrix Metalloproteinases 2 and 9

Journal: Journal of Clinical Neurology (Seoul, Korea)

doi: 10.3988/jcn.2011.7.2.69

Immunofluorescence staining illustrating the expressions of MMP-2 and MMP-9 produced by macrophages in carotid plaques. A and D: MMP-2 and MMP-9 were visualized with secondary antibodies conjugated to Texas Red (red). B and E: The nuclei were stained with DAPI (blue). C and F: MMPs colocalized with the cytoplasm in the macrophages. The white boxes in each image are magnified. Scale bar=100 µm. MMP: matrix metalloproteinase.
Figure Legend Snippet: Immunofluorescence staining illustrating the expressions of MMP-2 and MMP-9 produced by macrophages in carotid plaques. A and D: MMP-2 and MMP-9 were visualized with secondary antibodies conjugated to Texas Red (red). B and E: The nuclei were stained with DAPI (blue). C and F: MMPs colocalized with the cytoplasm in the macrophages. The white boxes in each image are magnified. Scale bar=100 µm. MMP: matrix metalloproteinase.

Techniques Used: Immunofluorescence, Staining, Produced

Carotid specimens that were immunohistochemically positive for MMP-2 (A) and MMP-9 (B) were also strongly positive for CD68 (C) but negative for SMA (D). MMP: matrix metalloproteinase, SMA: smooth-muscle actin.
Figure Legend Snippet: Carotid specimens that were immunohistochemically positive for MMP-2 (A) and MMP-9 (B) were also strongly positive for CD68 (C) but negative for SMA (D). MMP: matrix metalloproteinase, SMA: smooth-muscle actin.

Techniques Used:

62) Product Images from "Plaque Rupture is a Determinant of Vascular Events in Carotid Artery Atherosclerotic Disease: Involvement of Matrix Metalloproteinases 2 and 9"

Article Title: Plaque Rupture is a Determinant of Vascular Events in Carotid Artery Atherosclerotic Disease: Involvement of Matrix Metalloproteinases 2 and 9

Journal: Journal of Clinical Neurology (Seoul, Korea)

doi: 10.3988/jcn.2011.7.2.69

Immunofluorescence staining illustrating the expressions of MMP-2 and MMP-9 produced by macrophages in carotid plaques. A and D: MMP-2 and MMP-9 were visualized with secondary antibodies conjugated to Texas Red (red). B and E: The nuclei were stained with DAPI (blue). C and F: MMPs colocalized with the cytoplasm in the macrophages. The white boxes in each image are magnified. Scale bar=100 µm. MMP: matrix metalloproteinase.
Figure Legend Snippet: Immunofluorescence staining illustrating the expressions of MMP-2 and MMP-9 produced by macrophages in carotid plaques. A and D: MMP-2 and MMP-9 were visualized with secondary antibodies conjugated to Texas Red (red). B and E: The nuclei were stained with DAPI (blue). C and F: MMPs colocalized with the cytoplasm in the macrophages. The white boxes in each image are magnified. Scale bar=100 µm. MMP: matrix metalloproteinase.

Techniques Used: Immunofluorescence, Staining, Produced

Carotid specimens that were immunohistochemically positive for MMP-2 (A) and MMP-9 (B) were also strongly positive for CD68 (C) but negative for SMA (D). MMP: matrix metalloproteinase, SMA: smooth-muscle actin.
Figure Legend Snippet: Carotid specimens that were immunohistochemically positive for MMP-2 (A) and MMP-9 (B) were also strongly positive for CD68 (C) but negative for SMA (D). MMP: matrix metalloproteinase, SMA: smooth-muscle actin.

Techniques Used:

63) Product Images from "Plaque Rupture is a Determinant of Vascular Events in Carotid Artery Atherosclerotic Disease: Involvement of Matrix Metalloproteinases 2 and 9"

Article Title: Plaque Rupture is a Determinant of Vascular Events in Carotid Artery Atherosclerotic Disease: Involvement of Matrix Metalloproteinases 2 and 9

Journal: Journal of Clinical Neurology (Seoul, Korea)

doi: 10.3988/jcn.2011.7.2.69

Immunofluorescence staining illustrating the expressions of MMP-2 and MMP-9 produced by macrophages in carotid plaques. A and D: MMP-2 and MMP-9 were visualized with secondary antibodies conjugated to Texas Red (red). B and E: The nuclei were stained with DAPI (blue). C and F: MMPs colocalized with the cytoplasm in the macrophages. The white boxes in each image are magnified. Scale bar=100 µm. MMP: matrix metalloproteinase.
Figure Legend Snippet: Immunofluorescence staining illustrating the expressions of MMP-2 and MMP-9 produced by macrophages in carotid plaques. A and D: MMP-2 and MMP-9 were visualized with secondary antibodies conjugated to Texas Red (red). B and E: The nuclei were stained with DAPI (blue). C and F: MMPs colocalized with the cytoplasm in the macrophages. The white boxes in each image are magnified. Scale bar=100 µm. MMP: matrix metalloproteinase.

Techniques Used: Immunofluorescence, Staining, Produced

Carotid specimens that were immunohistochemically positive for MMP-2 (A) and MMP-9 (B) were also strongly positive for CD68 (C) but negative for SMA (D). MMP: matrix metalloproteinase, SMA: smooth-muscle actin.
Figure Legend Snippet: Carotid specimens that were immunohistochemically positive for MMP-2 (A) and MMP-9 (B) were also strongly positive for CD68 (C) but negative for SMA (D). MMP: matrix metalloproteinase, SMA: smooth-muscle actin.

Techniques Used:

64) Product Images from "Differential regulation of cell motility and invasion by FAK"

Article Title: Differential regulation of cell motility and invasion by FAK

Journal: The Journal of Cell Biology

doi: 10.1083/jcb.200212114

FAK and activated Q61L Rac synergize to promote JNK activation, increased MMP-9 expression, and an invasive cell phenotype. (A) Flag tag blotting was used to visualize Q61L Rac expression in the indicated Ad- infected cells. (B) Matrigel invasion assays were performed with the indicated Mock or Ad-Q61L Rac-infected cells. Values are means ± SD of triplicates from two independent experiments. (C) Gelatin zymography was performed with conditioned media from the indicated Mock or Ad-Q61L Rac-infected cells. Migration of pro and active forms of MMP-9 and MMP-2 are shown. (D) Conditioned media from FAK −/− v-Src or DA2 v-Src cells was concentrated, and MMP-9 or MMP-2 secretion was evaluated by blotting. (E) Semiquantitative RT-PCR was performed using primers to MMP-9 and to β-actin and showed that MMP-9 mRNA levels were 6.5-fold higher in DA2 v-Src compared with FAK −/− v-Src cells. (F) Matrigel invasion assays were performed with DA2 v-Src cells with the addition of recombinant TIMP-1 at the indicated concentration in the top chamber. Values are means ± SD of triplicates from two independent experiments. (G) JNK IP/IVK assays performed with lysates from FAK −/− cells or Ad-Q61L Rac-infected FAK −/− , FAK −/− v-Src, or DA2 v-Src cells. Values are means ± SD of duplicates from two independent experiments. Flag tag blotting of whole cell lysates was used to verify equivalent Q61L Rac expression.
Figure Legend Snippet: FAK and activated Q61L Rac synergize to promote JNK activation, increased MMP-9 expression, and an invasive cell phenotype. (A) Flag tag blotting was used to visualize Q61L Rac expression in the indicated Ad- infected cells. (B) Matrigel invasion assays were performed with the indicated Mock or Ad-Q61L Rac-infected cells. Values are means ± SD of triplicates from two independent experiments. (C) Gelatin zymography was performed with conditioned media from the indicated Mock or Ad-Q61L Rac-infected cells. Migration of pro and active forms of MMP-9 and MMP-2 are shown. (D) Conditioned media from FAK −/− v-Src or DA2 v-Src cells was concentrated, and MMP-9 or MMP-2 secretion was evaluated by blotting. (E) Semiquantitative RT-PCR was performed using primers to MMP-9 and to β-actin and showed that MMP-9 mRNA levels were 6.5-fold higher in DA2 v-Src compared with FAK −/− v-Src cells. (F) Matrigel invasion assays were performed with DA2 v-Src cells with the addition of recombinant TIMP-1 at the indicated concentration in the top chamber. Values are means ± SD of triplicates from two independent experiments. (G) JNK IP/IVK assays performed with lysates from FAK −/− cells or Ad-Q61L Rac-infected FAK −/− , FAK −/− v-Src, or DA2 v-Src cells. Values are means ± SD of duplicates from two independent experiments. Flag tag blotting of whole cell lysates was used to verify equivalent Q61L Rac expression.

Techniques Used: Activation Assay, Expressing, FLAG-tag, Infection, Zymography, Migration, Reverse Transcription Polymerase Chain Reaction, Recombinant, Concentration Assay

Models of differential FAK signaling connections at focal contacts or lamellipodia/invadopodia. (A) Integrin-associated FAK and Src activation promotes increased p190RhoGAP tyrosine phosphorylation or ERK2/MAP kinase activation. FAK–Src modulation of p21 Rho activity in FAK −/− cells is associated with increased cell motility, focal contact turnover, and actin cytoskeletal rearrangements. (B) In either EGF-stimulated human adenocarcinoma cells ( Hauck et al., 2001b ) or serum-stimulated v-Src–transformed fibroblasts, FAK functions to coordinate a Src-p130Cas signaling complex localized to lamellipodia in two-dimensional cell culture or to invadopodia in 3-D invasion assays. Through direct Cas SH3-mediated and indirect Crk-mediated interactions with the Dock180 guanine-nucleotide exchange factor, the FAK–Src signaling complex promotes Rac and JNK activation with effects upon gene expression. Although PI 3-kinase is a direct target of both v-Src and Rac ( Liu et al., 1993 ; Bishop and Hall, 2000 ), FAK −/− v-Src cells exhibit defects in JNK but not Akt activation. Overexpression of JNK in FAK −/− v-Src cells coupled with serum stimulation promoted c-Jun Ser-63 phosphorylation, MMP-9 expression, MMP-2 activation, and rescued the FAK −/− v-Src cell invasion defects.
Figure Legend Snippet: Models of differential FAK signaling connections at focal contacts or lamellipodia/invadopodia. (A) Integrin-associated FAK and Src activation promotes increased p190RhoGAP tyrosine phosphorylation or ERK2/MAP kinase activation. FAK–Src modulation of p21 Rho activity in FAK −/− cells is associated with increased cell motility, focal contact turnover, and actin cytoskeletal rearrangements. (B) In either EGF-stimulated human adenocarcinoma cells ( Hauck et al., 2001b ) or serum-stimulated v-Src–transformed fibroblasts, FAK functions to coordinate a Src-p130Cas signaling complex localized to lamellipodia in two-dimensional cell culture or to invadopodia in 3-D invasion assays. Through direct Cas SH3-mediated and indirect Crk-mediated interactions with the Dock180 guanine-nucleotide exchange factor, the FAK–Src signaling complex promotes Rac and JNK activation with effects upon gene expression. Although PI 3-kinase is a direct target of both v-Src and Rac ( Liu et al., 1993 ; Bishop and Hall, 2000 ), FAK −/− v-Src cells exhibit defects in JNK but not Akt activation. Overexpression of JNK in FAK −/− v-Src cells coupled with serum stimulation promoted c-Jun Ser-63 phosphorylation, MMP-9 expression, MMP-2 activation, and rescued the FAK −/− v-Src cell invasion defects.

Techniques Used: Activation Assay, Activity Assay, Transformation Assay, Cell Culture, Expressing, Over Expression

65) Product Images from "B7-H3 is Overexpressed in Patients Suffering Osteosarcoma and Associated with Tumor Aggressiveness and Metastasis"

Article Title: B7-H3 is Overexpressed in Patients Suffering Osteosarcoma and Associated with Tumor Aggressiveness and Metastasis

Journal: PLoS ONE

doi: 10.1371/journal.pone.0070689

Overexpression or silencing of B7-H3 expression regulates the expression of MMP-2 in osteosarcoma cells with western blot analysis. GAPDH was used as an internal control. Histogram represents densitometric analysis of the ratio of B7-H3, MMP-2 and GAPDH bands. Experiments were repeated at least 3 times and the mean value was calculated. * p
Figure Legend Snippet: Overexpression or silencing of B7-H3 expression regulates the expression of MMP-2 in osteosarcoma cells with western blot analysis. GAPDH was used as an internal control. Histogram represents densitometric analysis of the ratio of B7-H3, MMP-2 and GAPDH bands. Experiments were repeated at least 3 times and the mean value was calculated. * p

Techniques Used: Over Expression, Expressing, Western Blot

66) Product Images from "B7-H3 is Overexpressed in Patients Suffering Osteosarcoma and Associated with Tumor Aggressiveness and Metastasis"

Article Title: B7-H3 is Overexpressed in Patients Suffering Osteosarcoma and Associated with Tumor Aggressiveness and Metastasis

Journal: PLoS ONE

doi: 10.1371/journal.pone.0070689

Overexpression or silencing of B7-H3 expression regulates the expression of MMP-2 in osteosarcoma cells with western blot analysis. GAPDH was used as an internal control. Histogram represents densitometric analysis of the ratio of B7-H3, MMP-2 and GAPDH bands. Experiments were repeated at least 3 times and the mean value was calculated. * p
Figure Legend Snippet: Overexpression or silencing of B7-H3 expression regulates the expression of MMP-2 in osteosarcoma cells with western blot analysis. GAPDH was used as an internal control. Histogram represents densitometric analysis of the ratio of B7-H3, MMP-2 and GAPDH bands. Experiments were repeated at least 3 times and the mean value was calculated. * p

Techniques Used: Over Expression, Expressing, Western Blot

67) Product Images from "Lipopolysaccharide Induces Pro-Inflammatory Cytokines and MMP Production via TLR4 in Nasal Polyp-Derived Fibroblast and Organ Culture"

Article Title: Lipopolysaccharide Induces Pro-Inflammatory Cytokines and MMP Production via TLR4 in Nasal Polyp-Derived Fibroblast and Organ Culture

Journal: PLoS ONE

doi: 10.1371/journal.pone.0090683

Inhibitory effect of TLR4 antagonist on pro-inflammatory cytokines and MMP-1 production in nasal polyp organ cultures. Nasal polyp tissues were cultured and stimulated with LPS with or without LPS-RS. (A) Production of IL-6 and IL-8 was measured using ELISA. Protein expression level of MMP-1 was examined using western blotting (B) and density analysis (C). (D) MMP-1 secretion was measured using collagen zymography. Secretion of MMP-2 and MMP-9 was measured using gelatin zymography. (E) The expression pattern and localization of MMP-1 by immunohistochemical staining. Values are mean ± SEM of three independent samples. Significant difference from control (* P
Figure Legend Snippet: Inhibitory effect of TLR4 antagonist on pro-inflammatory cytokines and MMP-1 production in nasal polyp organ cultures. Nasal polyp tissues were cultured and stimulated with LPS with or without LPS-RS. (A) Production of IL-6 and IL-8 was measured using ELISA. Protein expression level of MMP-1 was examined using western blotting (B) and density analysis (C). (D) MMP-1 secretion was measured using collagen zymography. Secretion of MMP-2 and MMP-9 was measured using gelatin zymography. (E) The expression pattern and localization of MMP-1 by immunohistochemical staining. Values are mean ± SEM of three independent samples. Significant difference from control (* P

Techniques Used: Cell Culture, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Zymography, Immunohistochemistry, Staining

Effect of lipopolysaccharide on MMP expression in NPDFs. NPDFs were stimulated with LPS in the presence or absence of each MAPK inhibitor. Expression levels of MMP-1 , MMP-2 , and MMP-9 were determined using real-time PCR (A), western blotting (B) and density analysis (C). (D) MMP-1 secretion was measured using collagen zymography. MMP-2 and MMP-9 secretion levels were measured using gelatin zymography. (E) The effect of inhibiting the MAPK pathway on MMP-1 mRNA expression was examined using real-time PCR. Values are mean ± SEM of independent samples. * P
Figure Legend Snippet: Effect of lipopolysaccharide on MMP expression in NPDFs. NPDFs were stimulated with LPS in the presence or absence of each MAPK inhibitor. Expression levels of MMP-1 , MMP-2 , and MMP-9 were determined using real-time PCR (A), western blotting (B) and density analysis (C). (D) MMP-1 secretion was measured using collagen zymography. MMP-2 and MMP-9 secretion levels were measured using gelatin zymography. (E) The effect of inhibiting the MAPK pathway on MMP-1 mRNA expression was examined using real-time PCR. Values are mean ± SEM of independent samples. * P

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Zymography

68) Product Images from "Suppressions of Migration and Invasion by Cantharidin in TSGH-8301 Human Bladder Carcinoma Cells through the Inhibitions of Matrix Metalloproteinase-2/-9 Signaling"

Article Title: Suppressions of Migration and Invasion by Cantharidin in TSGH-8301 Human Bladder Carcinoma Cells through the Inhibitions of Matrix Metalloproteinase-2/-9 Signaling

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

doi: 10.1155/2013/190281

Cantharidin affects the levels of associated proteins and gene levels in migration and invasion of TSGH-8301 cells. Cells were treated with cantharidin (0, 1, and 2.5 μ M) for 24 h, and then cells were collected. The total protein extract was quantified and determined as described in Section 2 . The levels of p-p38, p-JNK1/2, p-ERK1/2, MMP-2, and MMP-9 protein expressions (a) were estimated by Western blotting as described in Section 2 . (b) The total RNA was extracted from cantharidin-treated cells, and the RNA samples were reverse-transcribed to cDNA for real-time PCR as described in Section 2 . The ratios between MMP-2, MMP-9, and GAPDH mRNA are used and data represents mean ± SD in duplicate of at least three independent experiments. * P
Figure Legend Snippet: Cantharidin affects the levels of associated proteins and gene levels in migration and invasion of TSGH-8301 cells. Cells were treated with cantharidin (0, 1, and 2.5 μ M) for 24 h, and then cells were collected. The total protein extract was quantified and determined as described in Section 2 . The levels of p-p38, p-JNK1/2, p-ERK1/2, MMP-2, and MMP-9 protein expressions (a) were estimated by Western blotting as described in Section 2 . (b) The total RNA was extracted from cantharidin-treated cells, and the RNA samples were reverse-transcribed to cDNA for real-time PCR as described in Section 2 . The ratios between MMP-2, MMP-9, and GAPDH mRNA are used and data represents mean ± SD in duplicate of at least three independent experiments. * P

Techniques Used: Migration, Western Blot, Real-time Polymerase Chain Reaction

Cantharidin suppresses the activities of matrix metalloproteinases (MMPs) in TSGH-8301 cells. Gelatin zymography was used to evaluate the activities of MMP-2 and MMP-9 as described in Section 2 . The different activity of MMP-2 and -9 was determined by densitometry analysis, and results are expressed as % of control. Similar results were obtained from three independent experiments.
Figure Legend Snippet: Cantharidin suppresses the activities of matrix metalloproteinases (MMPs) in TSGH-8301 cells. Gelatin zymography was used to evaluate the activities of MMP-2 and MMP-9 as described in Section 2 . The different activity of MMP-2 and -9 was determined by densitometry analysis, and results are expressed as % of control. Similar results were obtained from three independent experiments.

Techniques Used: Zymography, Activity Assay

69) Product Images from "Surgery-Induced Hippocampal Angiotensin II Elevation Causes Blood-Brain Barrier Disruption via MMP/TIMP in Aged Rats"

Article Title: Surgery-Induced Hippocampal Angiotensin II Elevation Causes Blood-Brain Barrier Disruption via MMP/TIMP in Aged Rats

Journal: Frontiers in Cellular Neuroscience

doi: 10.3389/fncel.2016.00105

Hippocampal matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) gene expression in response to surgery under isoflurane anesthesia. Twenty-month-old rats received laparotomy under isoflurane anesthesia. The levels of MMPs, including MMP-2 and MMP-9, as well as three of their endogenous tissue inhibitors (TIMP-1, -2, -3), were dynamically determined at 3, 6, 12, 24, and 72 h after surgery using qRT-PCR. Kinetics of surgery- induced changes in transcript levels of MMP-2 (A) , MMP-9 (B) , TIMPs (C) and MMP:TIMP ratios (D–I) in the hippocampus are shown. Values are mean ± SD, n = 6. * p
Figure Legend Snippet: Hippocampal matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) gene expression in response to surgery under isoflurane anesthesia. Twenty-month-old rats received laparotomy under isoflurane anesthesia. The levels of MMPs, including MMP-2 and MMP-9, as well as three of their endogenous tissue inhibitors (TIMP-1, -2, -3), were dynamically determined at 3, 6, 12, 24, and 72 h after surgery using qRT-PCR. Kinetics of surgery- induced changes in transcript levels of MMP-2 (A) , MMP-9 (B) , TIMPs (C) and MMP:TIMP ratios (D–I) in the hippocampus are shown. Values are mean ± SD, n = 6. * p

Techniques Used: Expressing, Quantitative RT-PCR

AT1 antagonist candesartan pretreatment attenuates surgery-induced imbalance between MMPs and TIMPs at the protein level. Twenty-month-old rats received laparotomy under isoflurane anesthesia in the presence or absence of candesartan pretreatment. Prior to surgery challenge, candesartan was intraperitoneally administered at a dose of 0.1 mg/kg daily for 14 consecutive days. (A) Representative bands of MMPs and TIMPs expression in the hippocampus at 6 h after surgery as detected by western blot analysis. (B) Semi-quantitative data showing protein expression levels of MMPs and TIMPs. Surgery upregulated MMP-9 protein expression and downregulated TIMP-3 expression. (C) The ratio of MMPs to TIMPs was calculated. The ratios of MMP-2/TIMP-3, MMP-9/TIMP-1, MMP-9/TIMP-2, and MMP-9/TIMP-3 were increased at 6 h post-surgery. These changes were all alleviated by candesartan treatment. Values are mean ± SD, n = 6. * p
Figure Legend Snippet: AT1 antagonist candesartan pretreatment attenuates surgery-induced imbalance between MMPs and TIMPs at the protein level. Twenty-month-old rats received laparotomy under isoflurane anesthesia in the presence or absence of candesartan pretreatment. Prior to surgery challenge, candesartan was intraperitoneally administered at a dose of 0.1 mg/kg daily for 14 consecutive days. (A) Representative bands of MMPs and TIMPs expression in the hippocampus at 6 h after surgery as detected by western blot analysis. (B) Semi-quantitative data showing protein expression levels of MMPs and TIMPs. Surgery upregulated MMP-9 protein expression and downregulated TIMP-3 expression. (C) The ratio of MMPs to TIMPs was calculated. The ratios of MMP-2/TIMP-3, MMP-9/TIMP-1, MMP-9/TIMP-2, and MMP-9/TIMP-3 were increased at 6 h post-surgery. These changes were all alleviated by candesartan treatment. Values are mean ± SD, n = 6. * p

Techniques Used: Expressing, Western Blot

70) Product Images from "Chrysin Inhibits Advanced Glycation End Products-Induced Kidney Fibrosis in Renal Mesangial Cells and Diabetic Kidneys"

Article Title: Chrysin Inhibits Advanced Glycation End Products-Induced Kidney Fibrosis in Renal Mesangial Cells and Diabetic Kidneys

Journal: Nutrients

doi: 10.3390/nu10070882

Suppressive effects of chrysin on cellular induction of α-SMA, fibroblast specific protein (FSP)-1, MM-2, MMP-9 and MT1-MMP in AGE-exposed HRMC. Renal mesangial cells were challenged for 3 days with 100 μg/mL AGE-BSA in the absence and presence of 1–20 μM chrysin. Cell extracts and media were subject to Western blot analysis with a primary antibody against α-SMA, FSP-1, MMP-2, MMP-9 or MT1-MMP ( A , D ). β-Actin protein was used as an internal control. The bar graphs in the right panel represent densitometric results obtained from Image analysis. The FSP-1 induction in AGE-exposed cells was immunocytochemically visualized as fluorescent Cy3 staining and nuclear counter-staining was done with the blue stain DAPI ( B , C ). Each photograph is representative of four different experiments and fluorescent images were taken with a fluorescence microscope. Scale bar = 100 µm. Fluorescent Cy3 staining intensity of FSP-1 was measured using an optical Axiomager microscope system ( B ). Values in bar graphs (mean ± SEM, n = 3 independent experiments) not sharing a same lower case indicate significant different at p
Figure Legend Snippet: Suppressive effects of chrysin on cellular induction of α-SMA, fibroblast specific protein (FSP)-1, MM-2, MMP-9 and MT1-MMP in AGE-exposed HRMC. Renal mesangial cells were challenged for 3 days with 100 μg/mL AGE-BSA in the absence and presence of 1–20 μM chrysin. Cell extracts and media were subject to Western blot analysis with a primary antibody against α-SMA, FSP-1, MMP-2, MMP-9 or MT1-MMP ( A , D ). β-Actin protein was used as an internal control. The bar graphs in the right panel represent densitometric results obtained from Image analysis. The FSP-1 induction in AGE-exposed cells was immunocytochemically visualized as fluorescent Cy3 staining and nuclear counter-staining was done with the blue stain DAPI ( B , C ). Each photograph is representative of four different experiments and fluorescent images were taken with a fluorescence microscope. Scale bar = 100 µm. Fluorescent Cy3 staining intensity of FSP-1 was measured using an optical Axiomager microscope system ( B ). Values in bar graphs (mean ± SEM, n = 3 independent experiments) not sharing a same lower case indicate significant different at p

Techniques Used: Western Blot, Staining, Fluorescence, Microscopy

71) Product Images from "FBP1 promotes ovarian cancer development through the acceleration of cell cycle transition and metastasis"

Article Title: FBP1 promotes ovarian cancer development through the acceleration of cell cycle transition and metastasis

Journal: Oncology Letters

doi: 10.3892/ol.2018.8872

The knockdown of FBP1 inhibited cell metastasis. (A) Knockdown of FBP1 inhibited cell migration analyzed by wound healing experiment. (B) Migratory distance of the wound-healing assay (C) Knockdown of FBP1 inhibited cell migration as analyzed by the Transwell assay. (D) MMP-2 expression in FBP1-C and FBP1-KD SKOV3 cells. Student's t-test was used to test statistical significance between groups. *P
Figure Legend Snippet: The knockdown of FBP1 inhibited cell metastasis. (A) Knockdown of FBP1 inhibited cell migration analyzed by wound healing experiment. (B) Migratory distance of the wound-healing assay (C) Knockdown of FBP1 inhibited cell migration as analyzed by the Transwell assay. (D) MMP-2 expression in FBP1-C and FBP1-KD SKOV3 cells. Student's t-test was used to test statistical significance between groups. *P

Techniques Used: Migration, Wound Healing Assay, Transwell Assay, Expressing

72) Product Images from "FBP1 promotes ovarian cancer development through the acceleration of cell cycle transition and metastasis"

Article Title: FBP1 promotes ovarian cancer development through the acceleration of cell cycle transition and metastasis

Journal: Oncology Letters

doi: 10.3892/ol.2018.8872

The knockdown of FBP1 inhibited cell metastasis. (A) Knockdown of FBP1 inhibited cell migration analyzed by wound healing experiment. (B) Migratory distance of the wound-healing assay (C) Knockdown of FBP1 inhibited cell migration as analyzed by the Transwell assay. (D) MMP-2 expression in FBP1-C and FBP1-KD SKOV3 cells. Student's t-test was used to test statistical significance between groups. *P
Figure Legend Snippet: The knockdown of FBP1 inhibited cell metastasis. (A) Knockdown of FBP1 inhibited cell migration analyzed by wound healing experiment. (B) Migratory distance of the wound-healing assay (C) Knockdown of FBP1 inhibited cell migration as analyzed by the Transwell assay. (D) MMP-2 expression in FBP1-C and FBP1-KD SKOV3 cells. Student's t-test was used to test statistical significance between groups. *P

Techniques Used: Migration, Wound Healing Assay, Transwell Assay, Expressing

73) Product Images from "Anti-proliferative effect of olmesartan on Tenon's capsule fibroblasts"

Article Title: Anti-proliferative effect of olmesartan on Tenon's capsule fibroblasts

Journal: International Journal of Ophthalmology

doi: 10.18240/ijo.2016.05.05

Olmesartan treated-eye presents with lower MMP-2 expression by means of mean density in each group comparing with the controlled eye.
Figure Legend Snippet: Olmesartan treated-eye presents with lower MMP-2 expression by means of mean density in each group comparing with the controlled eye.

Techniques Used: Expressing

The immunohistochemical pictures show the expression of MMP-2 in conjunctiva and subconjunctival tissue of 10×40 magnification
Figure Legend Snippet: The immunohistochemical pictures show the expression of MMP-2 in conjunctiva and subconjunctival tissue of 10×40 magnification

Techniques Used: Immunohistochemistry, Expressing

74) Product Images from "Anti-proliferative effect of olmesartan on Tenon's capsule fibroblasts"

Article Title: Anti-proliferative effect of olmesartan on Tenon's capsule fibroblasts

Journal: International Journal of Ophthalmology

doi: 10.18240/ijo.2016.05.05

Olmesartan treated-eye presents with lower MMP-2 expression by means of mean density in each group comparing with the controlled eye.
Figure Legend Snippet: Olmesartan treated-eye presents with lower MMP-2 expression by means of mean density in each group comparing with the controlled eye.

Techniques Used: Expressing

The immunohistochemical pictures show the expression of MMP-2 in conjunctiva and subconjunctival tissue of 10×40 magnification
Figure Legend Snippet: The immunohistochemical pictures show the expression of MMP-2 in conjunctiva and subconjunctival tissue of 10×40 magnification

Techniques Used: Immunohistochemistry, Expressing

75) Product Images from "Anti-proliferative effect of olmesartan on Tenon's capsule fibroblasts"

Article Title: Anti-proliferative effect of olmesartan on Tenon's capsule fibroblasts

Journal: International Journal of Ophthalmology

doi: 10.18240/ijo.2016.05.05

Olmesartan treated-eye presents with lower MMP-2 expression by means of mean density in each group comparing with the controlled eye.
Figure Legend Snippet: Olmesartan treated-eye presents with lower MMP-2 expression by means of mean density in each group comparing with the controlled eye.

Techniques Used: Expressing

The immunohistochemical pictures show the expression of MMP-2 in conjunctiva and subconjunctival tissue of 10×40 magnification
Figure Legend Snippet: The immunohistochemical pictures show the expression of MMP-2 in conjunctiva and subconjunctival tissue of 10×40 magnification

Techniques Used: Immunohistochemistry, Expressing

Related Articles

Luciferase:

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Article Snippet: .. Then we constructed colon cancer cells stably expressing luciferase as well as eIF4E, VEGF-C, MMP-2 and E-cadherin (HCT-15/RLuc/eIF4E, HCT-15/Rluc/VEGF-C, HCT-15/Rluc/MMP-2, and HCT-15/Rluc/E-cadherin). ..

Article Title: Rho GDIβ promotes Sp1/ MMP‐2 expression and bladder cancer invasion through perturbing miR‐200c‐targeted JNK2 protein translation
Article Snippet: The Sp1‐dependent luciferase reporter, which contains the three consensus binding sites of Sp1, has been described previously (Fang et al ., ). .. The antibodies for MMP‐2, RhoGDIβ, E2F1, HSF1, ETS‐1, p53, Smad4, Sp1, and β‐actin, were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Stable Transfection:

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Article Snippet: .. Then we constructed colon cancer cells stably expressing luciferase as well as eIF4E, VEGF-C, MMP-2 and E-cadherin (HCT-15/RLuc/eIF4E, HCT-15/Rluc/VEGF-C, HCT-15/Rluc/MMP-2, and HCT-15/Rluc/E-cadherin). ..

Immunostaining:

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Article Snippet: .. Immunohistochemistry The paraffin-embedded tissue sections were subjected to immunostaining with the use of polyclonal antibodies against eIF4E, VEGF-C, and MMP-2 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and mouse monoclonal antibodies against E-cadherin (Santa Cruz Biotechnology, Inc.). .. The primary antibody was diluted in phosphate-buffered saline with 1.5% normal blocking serum.

Enzyme-linked Immunosorbent Assay:

Article Title: MMP-2 Alters VEGF Expression via ?V?3 Integrin-Mediated PI3K/AKT Signaling in A549 Lung Cancer Cells
Article Snippet: We used antibodies specific for MMP-2, VEGF (VEGF-A), VEGFR-2, HIF-1α, GAPDH, integrin-αVβ3 (Clone 23C6), MT1MMP, (Santa Cruz Biotechnology, Santa Cruz, CA), PI3K, AKT, phospho-AKT (Ser-473) (Cell Signal Technology, Boston, MA), functional blocking integrin-αVβ3 (Clone 23C6; Cat# CBL544; Millipore Corporation, Temecula, CA) and anti-Human Von-Willebrand Factor (Factor-VIII; Dako North America, Inc, CA), and HRP/Alexa Fluor® conjugated secondary antibodies (Santa Cruz Biotechnology, Santa Cruz, CA). .. We also used constitutively active-AKT (myr-AKT) plasmid (Addgene, Plasmid 10841), human recombinant-MMP-2, human recombinant-VEGF165 (Millipore Corporation, Temecula, CA), human VEGF quantikine ELISA Kit, human MMP-2 quantikine ELISA Kit (R & D Systems, Minneapolis, MN) and ARP-100, a MMP-2 specific inhibitor (TOCRIS Bioscience, Ellisville, MO) in this study.

Microarray:

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells
Article Snippet: Immunohistochemical staining was carried out on tissue section from the tissue microarray (TMA). .. Then, sections were incubated with antibody against Aurora-A (Cell Signaling, Beverly, USA) or MMP-2 (Santa Cruz Biotechnology, Santa Cruz, USA) at 4°C for overnight, followed by incubation with biotinylated secondary antibody and streptavidin–peroxidase complex.

Incubation:

Article Title: MMP-2 Alters VEGF Expression via ?V?3 Integrin-Mediated PI3K/AKT Signaling in A549 Lung Cancer Cells
Article Snippet: Cells were incubated at 37 °C in a humidified 5 % CO2 atmosphere. .. We used antibodies specific for MMP-2, VEGF (VEGF-A), VEGFR-2, HIF-1α, GAPDH, integrin-αVβ3 (Clone 23C6), MT1MMP, (Santa Cruz Biotechnology, Santa Cruz, CA), PI3K, AKT, phospho-AKT (Ser-473) (Cell Signal Technology, Boston, MA), functional blocking integrin-αVβ3 (Clone 23C6; Cat# CBL544; Millipore Corporation, Temecula, CA) and anti-Human Von-Willebrand Factor (Factor-VIII; Dako North America, Inc, CA), and HRP/Alexa Fluor® conjugated secondary antibodies (Santa Cruz Biotechnology, Santa Cruz, CA).

Article Title: MMP-2 siRNA Inhibits Radiation-Enhanced Invasiveness in Glioma Cells
Article Snippet: After blocking with 5% nonfat dry milk and 0.1% Tween-20 in PBS, membranes were incubated with 1∶1000 dilution of primary antibodies followed by incubation in HRP-conjugated secondary antibodies. .. Primary antibodies used in this study were: MMP-2, MMP-9, VEGF, VEGFR-2, pVEGFR-2, pFAK, FAK, p-p38, total p38, caspase 3, caspase 8, caspase 9, GAPDH (used as a loading control), HRP conjugated secondary antibodies (Santa Cruz Biotechnology, SantaCruz, CA).

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells
Article Snippet: .. Then, sections were incubated with antibody against Aurora-A (Cell Signaling, Beverly, USA) or MMP-2 (Santa Cruz Biotechnology, Santa Cruz, USA) at 4°C for overnight, followed by incubation with biotinylated secondary antibody and streptavidin–peroxidase complex. .. After visualization of the reaction with the diaminobenzidine (DAB) chromogen, the slides were counterstained with haematoxylin.

Activity Assay:

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells
Article Snippet: Sections were deparaffinized in xylene, rehydrated in graded alcohol, immersed in 10 mM sodium citrate buffer (pH 6.0), and pretreated in a microwave oven for 10 min. Endogenous peroxidase activity was blocked with 0.3% hydrogen peroxide solution for 10 min. After being washed, the sections were treated with 1% bovine serum albumin for 30 min to block nonspecific reactions. .. Then, sections were incubated with antibody against Aurora-A (Cell Signaling, Beverly, USA) or MMP-2 (Santa Cruz Biotechnology, Santa Cruz, USA) at 4°C for overnight, followed by incubation with biotinylated secondary antibody and streptavidin–peroxidase complex.

Expressing:

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells
Article Snippet: .. Samantaray S, Sharma R, Chattopadhyaya TK, Gupta SD, Ralhan R. Increased expression of MMP-2 and MMP-9 in esophageal squamous cell carcinoma . .. J Cancer Res Clin Oncol 2004, 130 : 37–44.

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Article Snippet: .. Then we constructed colon cancer cells stably expressing luciferase as well as eIF4E, VEGF-C, MMP-2 and E-cadherin (HCT-15/RLuc/eIF4E, HCT-15/Rluc/VEGF-C, HCT-15/Rluc/MMP-2, and HCT-15/Rluc/E-cadherin). ..

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells
Article Snippet: .. The scores of < 5 were considered negative and the scores of ≥5 were considered positive for expression of Aurora-A or MMP-2 [ , ]. .. Whole cell extracts were prepared by using protein lysis buffer (150 mM NaCl, 1% Triton X-100, and 50 mM Tris–HCl, pH 8.0) supplemented with 1 mM phenylmethylsulfonyl fluoride, 5 μg/ml leupeptin, and 5 μg/ml aprotinin.

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells
Article Snippet: .. In conclusion, our results indicate that overexpression of Aurora-A is associated with the lymph node metastasis of ESCC and the high expression of MMP-2 in ESCC tissues. .. In addition, Aurora-A overexpression significantly increases cell invasion by upregulating MMP-2 expression in ESCC cells.

Article Title: Protection of the Transplant Kidney from Preservation Injury by Inhibition of Matrix Metalloproteinases
Article Snippet: .. Inhibition of MMP-2 with siRNA A stock solution of a mixture containing a pool of 3 target-specific 19–25 nucleotide small interfering RNAs designed to knock down rat MMP-2 gene expression (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used (10 μM concentration in a buffer containing 10 μM Tris-HCl, 20 mM NaCl, and 1 mM EDTA at pH 8.0). .. As a control, scrambled siRNA (Santa Cruz Biotechnology, Santa Cruz, CA) was used.

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells
Article Snippet: .. The results of western blot analysis ( Fig. A ) and qRT-PCR ( Fig. B ) showed that the protein and mRNA expression levels of MMP-2 in Aurora-A-overexpressing cells were higher than those in the control cells, which were consistent with previous studies that knockdown of Aurora-A expression could downregulate MMP-2 expression [ , ]. .. In addition, to verify that MMP-2 is required for Aurora-A-mediated cell invasion, Aurora-A-overexpressing cells were treated with MMP-2 inhibitor I.

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells
Article Snippet: .. An increased expression of MMP-2 has frequently been detected in solid tumors, which can contribute to cancer invasion and metastasis through the degradation of ECM, which facilitates and accelerates the tumor cells to invade new tissues and enter the blood stream to travel to distant sites [ – ]. .. In this study, we found that MMP-2 was overexpressed in ESCC, which was consistent with previous studies [ , ].

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells
Article Snippet: .. An investigation into the combined expression of Aurora-A and MMP-2 may be useful in the diagnosis and therapy of ESCC. .. As the expression of Aurora-A protein was found to be correlated with MMP-2 expression in the patient cohort, we further analyzed their expressions in ESCC cell lines ( Fig. ).

Western Blot:

Article Title: MMP-2 siRNA Inhibits Radiation-Enhanced Invasiveness in Glioma Cells
Article Snippet: Paragraph title: Western blot analysis ... Primary antibodies used in this study were: MMP-2, MMP-9, VEGF, VEGFR-2, pVEGFR-2, pFAK, FAK, p-p38, total p38, caspase 3, caspase 8, caspase 9, GAPDH (used as a loading control), HRP conjugated secondary antibodies (Santa Cruz Biotechnology, SantaCruz, CA).

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells
Article Snippet: .. The results of western blot analysis ( Fig. A ) and qRT-PCR ( Fig. B ) showed that the protein and mRNA expression levels of MMP-2 in Aurora-A-overexpressing cells were higher than those in the control cells, which were consistent with previous studies that knockdown of Aurora-A expression could downregulate MMP-2 expression [ , ]. .. In addition, to verify that MMP-2 is required for Aurora-A-mediated cell invasion, Aurora-A-overexpressing cells were treated with MMP-2 inhibitor I.

Over Expression:

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells
Article Snippet: .. In conclusion, our results indicate that overexpression of Aurora-A is associated with the lymph node metastasis of ESCC and the high expression of MMP-2 in ESCC tissues. .. In addition, Aurora-A overexpression significantly increases cell invasion by upregulating MMP-2 expression in ESCC cells.

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells
Article Snippet: .. Li Y, Ma J, Guo Q, Duan F, Tang F, Zheng P, Zhao Z, et al. Overexpression of MMP-2 and MMP-9 in esophageal squamous cell carcinoma . .. Dis Esophagus 2009, 22 : 664–667.

Blocking Assay:

Article Title: Expression analysis and clinical significance of eIF4E, VEGF-C, E-cadherin and MMP-2 in colorectal adenocarcinoma
Article Snippet: Immunohistochemistry The paraffin-embedded tissue sections were subjected to immunostaining with the use of polyclonal antibodies against eIF4E, VEGF-C, and MMP-2 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and mouse monoclonal antibodies against E-cadherin (Santa Cruz Biotechnology, Inc.). .. The primary antibody was diluted in phosphate-buffered saline with 1.5% normal blocking serum.

Article Title: MMP-2 Alters VEGF Expression via ?V?3 Integrin-Mediated PI3K/AKT Signaling in A549 Lung Cancer Cells
Article Snippet: .. We used antibodies specific for MMP-2, VEGF (VEGF-A), VEGFR-2, HIF-1α, GAPDH, integrin-αVβ3 (Clone 23C6), MT1MMP, (Santa Cruz Biotechnology, Santa Cruz, CA), PI3K, AKT, phospho-AKT (Ser-473) (Cell Signal Technology, Boston, MA), functional blocking integrin-αVβ3 (Clone 23C6; Cat# CBL544; Millipore Corporation, Temecula, CA) and anti-Human Von-Willebrand Factor (Factor-VIII; Dako North America, Inc, CA), and HRP/Alexa Fluor® conjugated secondary antibodies (Santa Cruz Biotechnology, Santa Cruz, CA). .. We also used constitutively active-AKT (myr-AKT) plasmid (Addgene, Plasmid 10841), human recombinant-MMP-2, human recombinant-VEGF165 (Millipore Corporation, Temecula, CA), human VEGF quantikine ELISA Kit, human MMP-2 quantikine ELISA Kit (R & D Systems, Minneapolis, MN) and ARP-100, a MMP-2 specific inhibitor (TOCRIS Bioscience, Ellisville, MO) in this study.

Article Title: MMP-2 siRNA Inhibits Radiation-Enhanced Invasiveness in Glioma Cells
Article Snippet: After blocking with 5% nonfat dry milk and 0.1% Tween-20 in PBS, membranes were incubated with 1∶1000 dilution of primary antibodies followed by incubation in HRP-conjugated secondary antibodies. .. Primary antibodies used in this study were: MMP-2, MMP-9, VEGF, VEGFR-2, pVEGFR-2, pFAK, FAK, p-p38, total p38, caspase 3, caspase 8, caspase 9, GAPDH (used as a loading control), HRP conjugated secondary antibodies (Santa Cruz Biotechnology, SantaCruz, CA).

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells
Article Snippet: Sections were deparaffinized in xylene, rehydrated in graded alcohol, immersed in 10 mM sodium citrate buffer (pH 6.0), and pretreated in a microwave oven for 10 min. Endogenous peroxidase activity was blocked with 0.3% hydrogen peroxide solution for 10 min. After being washed, the sections were treated with 1% bovine serum albumin for 30 min to block nonspecific reactions. .. Then, sections were incubated with antibody against Aurora-A (Cell Signaling, Beverly, USA) or MMP-2 (Santa Cruz Biotechnology, Santa Cruz, USA) at 4°C for overnight, followed by incubation with biotinylated secondary antibody and streptavidin–peroxidase complex.

Countercurrent Chromatography:

Article Title: Rho GDIβ promotes Sp1/ MMP‐2 expression and bladder cancer invasion through perturbing miR‐200c‐targeted JNK2 protein translation
Article Snippet: The mutation of mmp‐2 promoter‐driven luciferase at the Sp1 binding site was made by point mutation using the following primers: Sense: 5′‐GTA GGG GGG TGG GGC AGA GAG ATA CGG GCC CGA GTG CGC CC‐3′ anti‐sense: 5′‐GGG CGC ACT CGG GCC CGT ATC TCT CTG CCC CAC CCC CCT AC‐3′. .. The antibodies for MMP‐2, RhoGDIβ, E2F1, HSF1, ETS‐1, p53, Smad4, Sp1, and β‐actin, were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Immunohistochemistry:

Article Title: Expression analysis and clinical significance of eIF4E, VEGF-C, E-cadherin and MMP-2 in colorectal adenocarcinoma
Article Snippet: .. Immunohistochemistry The paraffin-embedded tissue sections were subjected to immunostaining with the use of polyclonal antibodies against eIF4E, VEGF-C, and MMP-2 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and mouse monoclonal antibodies against E-cadherin (Santa Cruz Biotechnology, Inc.). .. The primary antibody was diluted in phosphate-buffered saline with 1.5% normal blocking serum.

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells
Article Snippet: Paragraph title: Immunohistochemical staining ... Then, sections were incubated with antibody against Aurora-A (Cell Signaling, Beverly, USA) or MMP-2 (Santa Cruz Biotechnology, Santa Cruz, USA) at 4°C for overnight, followed by incubation with biotinylated secondary antibody and streptavidin–peroxidase complex.

Cell Culture:

Article Title: MMP-2 Alters VEGF Expression via ?V?3 Integrin-Mediated PI3K/AKT Signaling in A549 Lung Cancer Cells
Article Snippet: A549 and H1299 lung adenocarcinoma cells were cultured in RPMI 1640 (ATCC Manassas, VA) supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, CA), 50 units/mL penicillin, and 50 μg/mL streptomycin (Life Technologies, Inc., Frederick, MD). .. We used antibodies specific for MMP-2, VEGF (VEGF-A), VEGFR-2, HIF-1α, GAPDH, integrin-αVβ3 (Clone 23C6), MT1MMP, (Santa Cruz Biotechnology, Santa Cruz, CA), PI3K, AKT, phospho-AKT (Ser-473) (Cell Signal Technology, Boston, MA), functional blocking integrin-αVβ3 (Clone 23C6; Cat# CBL544; Millipore Corporation, Temecula, CA) and anti-Human Von-Willebrand Factor (Factor-VIII; Dako North America, Inc, CA), and HRP/Alexa Fluor® conjugated secondary antibodies (Santa Cruz Biotechnology, Santa Cruz, CA).

Article Title: Rho GDIβ promotes Sp1/ MMP‐2 expression and bladder cancer invasion through perturbing miR‐200c‐targeted JNK2 protein translation
Article Snippet: Both cell lines were cultured in DMEM:F‐12 1 : 1 with 10% fetal bovine serum (FBS; ATLANTA, Flowery Branch, GA, USA) as described in our previous studies (Jiang et al ., ; Jin et al ., ), and cell lines will be authenticated every 6–12 months by Genetica DNA Laboratories (Burlington, NC 27215, CA, USA) and the results compared with the data in the ATCC STR database. .. The antibodies for MMP‐2, RhoGDIβ, E2F1, HSF1, ETS‐1, p53, Smad4, Sp1, and β‐actin, were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Inhibition:

Article Title: Protection of the Transplant Kidney from Preservation Injury by Inhibition of Matrix Metalloproteinases
Article Snippet: .. Inhibition of MMP-2 with siRNA A stock solution of a mixture containing a pool of 3 target-specific 19–25 nucleotide small interfering RNAs designed to knock down rat MMP-2 gene expression (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used (10 μM concentration in a buffer containing 10 μM Tris-HCl, 20 mM NaCl, and 1 mM EDTA at pH 8.0). .. As a control, scrambled siRNA (Santa Cruz Biotechnology, Santa Cruz, CA) was used.

Quantitative RT-PCR:

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells
Article Snippet: .. The results of western blot analysis ( Fig. A ) and qRT-PCR ( Fig. B ) showed that the protein and mRNA expression levels of MMP-2 in Aurora-A-overexpressing cells were higher than those in the control cells, which were consistent with previous studies that knockdown of Aurora-A expression could downregulate MMP-2 expression [ , ]. .. In addition, to verify that MMP-2 is required for Aurora-A-mediated cell invasion, Aurora-A-overexpressing cells were treated with MMP-2 inhibitor I.

Recombinant:

Article Title: MMP-2 Alters VEGF Expression via ?V?3 Integrin-Mediated PI3K/AKT Signaling in A549 Lung Cancer Cells
Article Snippet: We used antibodies specific for MMP-2, VEGF (VEGF-A), VEGFR-2, HIF-1α, GAPDH, integrin-αVβ3 (Clone 23C6), MT1MMP, (Santa Cruz Biotechnology, Santa Cruz, CA), PI3K, AKT, phospho-AKT (Ser-473) (Cell Signal Technology, Boston, MA), functional blocking integrin-αVβ3 (Clone 23C6; Cat# CBL544; Millipore Corporation, Temecula, CA) and anti-Human Von-Willebrand Factor (Factor-VIII; Dako North America, Inc, CA), and HRP/Alexa Fluor® conjugated secondary antibodies (Santa Cruz Biotechnology, Santa Cruz, CA). .. We also used constitutively active-AKT (myr-AKT) plasmid (Addgene, Plasmid 10841), human recombinant-MMP-2, human recombinant-VEGF165 (Millipore Corporation, Temecula, CA), human VEGF quantikine ELISA Kit, human MMP-2 quantikine ELISA Kit (R & D Systems, Minneapolis, MN) and ARP-100, a MMP-2 specific inhibitor (TOCRIS Bioscience, Ellisville, MO) in this study.

Mutagenesis:

Article Title: Rho GDIβ promotes Sp1/ MMP‐2 expression and bladder cancer invasion through perturbing miR‐200c‐targeted JNK2 protein translation
Article Snippet: The mutation of mmp‐2 promoter‐driven luciferase at the Sp1 binding site was made by point mutation using the following primers: Sense: 5′‐GTA GGG GGG TGG GGC AGA GAG ATA CGG GCC CGA GTG CGC CC‐3′ anti‐sense: 5′‐GGG CGC ACT CGG GCC CGT ATC TCT CTG CCC CAC CCC CCT AC‐3′. .. The antibodies for MMP‐2, RhoGDIβ, E2F1, HSF1, ETS‐1, p53, Smad4, Sp1, and β‐actin, were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Transfection:

Article Title: MMP-2 siRNA Inhibits Radiation-Enhanced Invasiveness in Glioma Cells
Article Snippet: Western blot analysis U-251 and U-87 cells were transfected and irradiated as described earlier. .. Primary antibodies used in this study were: MMP-2, MMP-9, VEGF, VEGFR-2, pVEGFR-2, pFAK, FAK, p-p38, total p38, caspase 3, caspase 8, caspase 9, GAPDH (used as a loading control), HRP conjugated secondary antibodies (Santa Cruz Biotechnology, SantaCruz, CA).

shRNA:

Article Title: Rho GDIβ promotes Sp1/ MMP‐2 expression and bladder cancer invasion through perturbing miR‐200c‐targeted JNK2 protein translation
Article Snippet: The short hairpin RNA constructs specific to Sp1(shSp1), JNK2(shJNK2), MMP‐2(shMMP‐2), and their scramble nonsense control construct were purchased from Open Biosystem (Pittsburgh, PA, USA). .. The antibodies for MMP‐2, RhoGDIβ, E2F1, HSF1, ETS‐1, p53, Smad4, Sp1, and β‐actin, were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Construct:

Article Title: Expression analysis and clinical significance of eIF4E, VEGF-C, E-cadherin and MMP-2 in colorectal adenocarcinoma
Article Snippet: .. Then we constructed colon cancer cells stably expressing luciferase as well as eIF4E, VEGF-C, MMP-2 and E-cadherin (HCT-15/RLuc/eIF4E, HCT-15/Rluc/VEGF-C, HCT-15/Rluc/MMP-2, and HCT-15/Rluc/E-cadherin). ..

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells
Article Snippet: TMA was constructed with a Beecher Instruments Tissue Array from 70 ESCC and paired normal adjacent tissue samples, which were obtained from the National Engineering Center for Biochip (Shanghai, China). .. Then, sections were incubated with antibody against Aurora-A (Cell Signaling, Beverly, USA) or MMP-2 (Santa Cruz Biotechnology, Santa Cruz, USA) at 4°C for overnight, followed by incubation with biotinylated secondary antibody and streptavidin–peroxidase complex.

Article Title: Rho GDIβ promotes Sp1/ MMP‐2 expression and bladder cancer invasion through perturbing miR‐200c‐targeted JNK2 protein translation
Article Snippet: The short hairpin RNA constructs specific to Sp1(shSp1), JNK2(shJNK2), MMP‐2(shMMP‐2), and their scramble nonsense control construct were purchased from Open Biosystem (Pittsburgh, PA, USA). .. The antibodies for MMP‐2, RhoGDIβ, E2F1, HSF1, ETS‐1, p53, Smad4, Sp1, and β‐actin, were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Activated Clotting Time Assay:

Article Title: Rho GDIβ promotes Sp1/ MMP‐2 expression and bladder cancer invasion through perturbing miR‐200c‐targeted JNK2 protein translation
Article Snippet: The mutation of mmp‐2 promoter‐driven luciferase at the Sp1 binding site was made by point mutation using the following primers: Sense: 5′‐GTA GGG GGG TGG GGC AGA GAG ATA CGG GCC CGA GTG CGC CC‐3′ anti‐sense: 5′‐GGG CGC ACT CGG GCC CGT ATC TCT CTG CCC CAC CCC CCT AC‐3′. .. The antibodies for MMP‐2, RhoGDIβ, E2F1, HSF1, ETS‐1, p53, Smad4, Sp1, and β‐actin, were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

SDS Page:

Article Title: MMP-2 siRNA Inhibits Radiation-Enhanced Invasiveness in Glioma Cells
Article Snippet: Equal amounts of total protein from cell lysates obtained by lysing cells in a suitable buffer [50 mM/L Tris-HCl (pH 7.4), 150 mM/L NaCl, 1% IGEPAL, 1 mM/L EDTA, 0.25% sodium deoxycholate, 1 mM/L sodium fluoride, 1 mM/L sodium orthovanadate, 0.5 mM/L PMSF, 10 µg/mL aprotinin, 10 µg/mL leupeptin] were separated by SDS–PAGE and transferred to polyvinylidene difluoride membranes (Bio-Rad, Hercules, CA). .. Primary antibodies used in this study were: MMP-2, MMP-9, VEGF, VEGFR-2, pVEGFR-2, pFAK, FAK, p-p38, total p38, caspase 3, caspase 8, caspase 9, GAPDH (used as a loading control), HRP conjugated secondary antibodies (Santa Cruz Biotechnology, SantaCruz, CA).

Plasmid Preparation:

Article Title: MMP-2 Alters VEGF Expression via ?V?3 Integrin-Mediated PI3K/AKT Signaling in A549 Lung Cancer Cells
Article Snippet: We used antibodies specific for MMP-2, VEGF (VEGF-A), VEGFR-2, HIF-1α, GAPDH, integrin-αVβ3 (Clone 23C6), MT1MMP, (Santa Cruz Biotechnology, Santa Cruz, CA), PI3K, AKT, phospho-AKT (Ser-473) (Cell Signal Technology, Boston, MA), functional blocking integrin-αVβ3 (Clone 23C6; Cat# CBL544; Millipore Corporation, Temecula, CA) and anti-Human Von-Willebrand Factor (Factor-VIII; Dako North America, Inc, CA), and HRP/Alexa Fluor® conjugated secondary antibodies (Santa Cruz Biotechnology, Santa Cruz, CA). .. We also used constitutively active-AKT (myr-AKT) plasmid (Addgene, Plasmid 10841), human recombinant-MMP-2, human recombinant-VEGF165 (Millipore Corporation, Temecula, CA), human VEGF quantikine ELISA Kit, human MMP-2 quantikine ELISA Kit (R & D Systems, Minneapolis, MN) and ARP-100, a MMP-2 specific inhibitor (TOCRIS Bioscience, Ellisville, MO) in this study.

Irradiation:

Article Title: MMP-2 siRNA Inhibits Radiation-Enhanced Invasiveness in Glioma Cells
Article Snippet: Western blot analysis U-251 and U-87 cells were transfected and irradiated as described earlier. .. Primary antibodies used in this study were: MMP-2, MMP-9, VEGF, VEGFR-2, pVEGFR-2, pFAK, FAK, p-p38, total p38, caspase 3, caspase 8, caspase 9, GAPDH (used as a loading control), HRP conjugated secondary antibodies (Santa Cruz Biotechnology, SantaCruz, CA).

Functional Assay:

Article Title: MMP-2 Alters VEGF Expression via ?V?3 Integrin-Mediated PI3K/AKT Signaling in A549 Lung Cancer Cells
Article Snippet: .. We used antibodies specific for MMP-2, VEGF (VEGF-A), VEGFR-2, HIF-1α, GAPDH, integrin-αVβ3 (Clone 23C6), MT1MMP, (Santa Cruz Biotechnology, Santa Cruz, CA), PI3K, AKT, phospho-AKT (Ser-473) (Cell Signal Technology, Boston, MA), functional blocking integrin-αVβ3 (Clone 23C6; Cat# CBL544; Millipore Corporation, Temecula, CA) and anti-Human Von-Willebrand Factor (Factor-VIII; Dako North America, Inc, CA), and HRP/Alexa Fluor® conjugated secondary antibodies (Santa Cruz Biotechnology, Santa Cruz, CA). .. We also used constitutively active-AKT (myr-AKT) plasmid (Addgene, Plasmid 10841), human recombinant-MMP-2, human recombinant-VEGF165 (Millipore Corporation, Temecula, CA), human VEGF quantikine ELISA Kit, human MMP-2 quantikine ELISA Kit (R & D Systems, Minneapolis, MN) and ARP-100, a MMP-2 specific inhibitor (TOCRIS Bioscience, Ellisville, MO) in this study.

Binding Assay:

Article Title: Rho GDIβ promotes Sp1/ MMP‐2 expression and bladder cancer invasion through perturbing miR‐200c‐targeted JNK2 protein translation
Article Snippet: The Sp1‐dependent luciferase reporter, which contains the three consensus binding sites of Sp1, has been described previously (Fang et al ., ). .. The antibodies for MMP‐2, RhoGDIβ, E2F1, HSF1, ETS‐1, p53, Smad4, Sp1, and β‐actin, were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Concentration Assay:

Article Title: Protection of the Transplant Kidney from Preservation Injury by Inhibition of Matrix Metalloproteinases
Article Snippet: .. Inhibition of MMP-2 with siRNA A stock solution of a mixture containing a pool of 3 target-specific 19–25 nucleotide small interfering RNAs designed to knock down rat MMP-2 gene expression (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used (10 μM concentration in a buffer containing 10 μM Tris-HCl, 20 mM NaCl, and 1 mM EDTA at pH 8.0). .. As a control, scrambled siRNA (Santa Cruz Biotechnology, Santa Cruz, CA) was used.

CTG Assay:

Article Title: Rho GDIβ promotes Sp1/ MMP‐2 expression and bladder cancer invasion through perturbing miR‐200c‐targeted JNK2 protein translation
Article Snippet: The mutation of mmp‐2 promoter‐driven luciferase at the Sp1 binding site was made by point mutation using the following primers: Sense: 5′‐GTA GGG GGG TGG GGC AGA GAG ATA CGG GCC CGA GTG CGC CC‐3′ anti‐sense: 5′‐GGG CGC ACT CGG GCC CGT ATC TCT CTG CCC CAC CCC CCT AC‐3′. .. The antibodies for MMP‐2, RhoGDIβ, E2F1, HSF1, ETS‐1, p53, Smad4, Sp1, and β‐actin, were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Staining:

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells
Article Snippet: Paragraph title: Immunohistochemical staining ... Then, sections were incubated with antibody against Aurora-A (Cell Signaling, Beverly, USA) or MMP-2 (Santa Cruz Biotechnology, Santa Cruz, USA) at 4°C for overnight, followed by incubation with biotinylated secondary antibody and streptavidin–peroxidase complex.

Article Title: Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells
Article Snippet: .. Additionally, negative cytoplasmic and weakly nuclear staining of MMP-2 was observed in adjacent normal tissues ( Fig. A ). .. In contrast, MMP-2 cytoplasmic staining was upregulated in ESCC tissues ( Fig. B–D ).

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  • 79
    Santa Cruz Biotechnology rat mmp 2 gene expression
    Comparison of MMP-9 and <t>MMP-2</t> levels in perfusates from kidneys from DBD (n = 15) and cDCDD (n = 9) human donors. Error bars represent Standard Error of the Mean (SEM). * p
    Rat Mmp 2 Gene Expression, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 79/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Santa Cruz Biotechnology mmp 2
    VEGF promotes increased <t>MMP-2</t> activity by increasing MT1-MMP expression in ECs. A : MT1-MMP and TIMP-2 mRNA levels, normalized to β-actin mRNA after exposure of iHUVECs to hypoxia for 0–8 h, reported as fold induction compared with untreated controls. B : MT1-MMP and TIMP-2 mRNA levels, normalized to β-actin mRNA after treatment of iHUVECs with increasing doses of rhVEGF for 6 h or increasing duration of treatment with 10 ng rhVEGF, reported as fold induction compared with untreated controls. C : Western blot for MT1-MMP in iHUVECs treated with increasing doses of rhVEGF (for 8 h) or increasing duration of treatment (with 10 ng rhVEGF). GAPDH was used as a loading control. D : MT1-MMP and TIMP-2 mRNA levels (RT-PCR) in iHUVECs treated with conditioned media from MIO-M1 cells exposed to hypoxia (0–6 h) with or without RNAi against VEGF mRNA reported as fold induction compared with untreated controls. E : MMP-2 enzymatic (gelatinase) activity after treatment of iHUVECs with increasing doses of rhVEGF (for 6 h) or increasing duration of treatment (with 10 ng rhVEGF), reported as fold induction compared with untreated controls. All experiments were performed in duplicate and are representative of at least three independent experiments. n = 3 animals in each group. Student t test: * P
    Mmp 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mmp 2/product/Santa Cruz Biotechnology
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    96
    Santa Cruz Biotechnology anti mmp 2
    <t>p-MMP-2</t> combined with radiation enhances apoptosis in vivo . A, Immunohistochemical analysis of brain sections using anti-MMP-2, anti-VEGF and anti-pFAK antibodies. Sections were photographed (60×). Also shown is the negative control where the primary antibody was replaced by non-specific IgG (insets). B, Tissue sections of mice were evaluated with the TUNEL assay according to manufacturer's instructions and photographed under fluorescent microscopy (60×). For the negative control, samples were incubated with label solution (without terminal transferase) instead of TUNEL reaction mixture (insets). C, siRNA against MMP-2 inhibits U251 tumor cell invasion in vivo . H E staining was performed according to standard protocol, and representative pictures of tumor sections from mock, pSV, p-MMP-2-treated mice are shown (20× and 60×). D , Immunohistochemical analysis of brain sections using anti-human nuclei (HuNu) antibody, a histological marker for identification of human cells (a specific human nuclear antigen). Entire brain sections were photographed (4×; middle row); shown on the top row is a non-tumor region (40×; top row); and shown on the bottom row is tumor and non-tumor overlapping region (40×; bottom row). Also shown is the negative control where the primary antibody was replaced by non-specific IgG (inset).
    Anti Mmp 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 56 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti mmp 2/product/Santa Cruz Biotechnology
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    Image Search Results


    Comparison of MMP-9 and MMP-2 levels in perfusates from kidneys from DBD (n = 15) and cDCDD (n = 9) human donors. Error bars represent Standard Error of the Mean (SEM). * p

    Journal: PLoS ONE

    Article Title: Protection of the Transplant Kidney from Preservation Injury by Inhibition of Matrix Metalloproteinases

    doi: 10.1371/journal.pone.0157508

    Figure Lengend Snippet: Comparison of MMP-9 and MMP-2 levels in perfusates from kidneys from DBD (n = 15) and cDCDD (n = 9) human donors. Error bars represent Standard Error of the Mean (SEM). * p

    Article Snippet: Inhibition of MMP-2 with siRNA A stock solution of a mixture containing a pool of 3 target-specific 19–25 nucleotide small interfering RNAs designed to knock down rat MMP-2 gene expression (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used (10 μM concentration in a buffer containing 10 μM Tris-HCl, 20 mM NaCl, and 1 mM EDTA at pH 8.0).

    Techniques:

    MMP-2 expression in perfusate from rat kidney cold perfused with KPS-1 solution with MMP-2 siRNA added (n = 4). Insert: a representative zymography for MMP-2. Error bars represent Standard Error of the Mean (SEM). * p

    Journal: PLoS ONE

    Article Title: Protection of the Transplant Kidney from Preservation Injury by Inhibition of Matrix Metalloproteinases

    doi: 10.1371/journal.pone.0157508

    Figure Lengend Snippet: MMP-2 expression in perfusate from rat kidney cold perfused with KPS-1 solution with MMP-2 siRNA added (n = 4). Insert: a representative zymography for MMP-2. Error bars represent Standard Error of the Mean (SEM). * p

    Article Snippet: Inhibition of MMP-2 with siRNA A stock solution of a mixture containing a pool of 3 target-specific 19–25 nucleotide small interfering RNAs designed to knock down rat MMP-2 gene expression (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used (10 μM concentration in a buffer containing 10 μM Tris-HCl, 20 mM NaCl, and 1 mM EDTA at pH 8.0).

    Techniques: Expressing, Zymography

    Level of undamaged mitochondria measured by the levels of cytochrome c oxidase activity in rat kidney tissue after initial flush and following 22 hours of machine cold perfusion with KPS-1, KPS-1 with doxycycline, and KPS-1 with MMP-2 siRNA (n = 4). Error bars represent Standard Error of the Mean (SEM). * p

    Journal: PLoS ONE

    Article Title: Protection of the Transplant Kidney from Preservation Injury by Inhibition of Matrix Metalloproteinases

    doi: 10.1371/journal.pone.0157508

    Figure Lengend Snippet: Level of undamaged mitochondria measured by the levels of cytochrome c oxidase activity in rat kidney tissue after initial flush and following 22 hours of machine cold perfusion with KPS-1, KPS-1 with doxycycline, and KPS-1 with MMP-2 siRNA (n = 4). Error bars represent Standard Error of the Mean (SEM). * p

    Article Snippet: Inhibition of MMP-2 with siRNA A stock solution of a mixture containing a pool of 3 target-specific 19–25 nucleotide small interfering RNAs designed to knock down rat MMP-2 gene expression (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used (10 μM concentration in a buffer containing 10 μM Tris-HCl, 20 mM NaCl, and 1 mM EDTA at pH 8.0).

    Techniques: Activity Assay

    Matrix Metalloproteinase levels and injury markers in perfusate from clinically transplanted kidneys with DGF (n = 5) and without DGF (n = 19). A: MMP-9 level B: MMP-2 level C: NGAL levelD: LDH activity Error bars represent Standard Error of the Mean (SEM). * p

    Journal: PLoS ONE

    Article Title: Protection of the Transplant Kidney from Preservation Injury by Inhibition of Matrix Metalloproteinases

    doi: 10.1371/journal.pone.0157508

    Figure Lengend Snippet: Matrix Metalloproteinase levels and injury markers in perfusate from clinically transplanted kidneys with DGF (n = 5) and without DGF (n = 19). A: MMP-9 level B: MMP-2 level C: NGAL levelD: LDH activity Error bars represent Standard Error of the Mean (SEM). * p

    Article Snippet: Inhibition of MMP-2 with siRNA A stock solution of a mixture containing a pool of 3 target-specific 19–25 nucleotide small interfering RNAs designed to knock down rat MMP-2 gene expression (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used (10 μM concentration in a buffer containing 10 μM Tris-HCl, 20 mM NaCl, and 1 mM EDTA at pH 8.0).

    Techniques: Activity Assay

    Effect of doxycycline on LDH, cytochrome c oxidase, and total protein release. A: Confirmation of inhibition of MMP-2 and MMP-9 activity by doxycycline using zymography. B: LDH activity in perfusate of rat kidney with warm ischemic injury at 5, 10, and 22 hours perfused with KPS-1 with and without doxycycline (n = 7 each). C: Cytochrome c oxidase level in perfusate of rat kidney with warm ischemic injury at 5, 10, and 22 hours perfused with KPS-1 with and without doxycycline (n = 7 each). D: Total protein release in perfusate of rat kidney with warm ischemic injury at 5, 10, and 22 hours perfused with KPS-1 with and without doxycycline (n = 6 each). Error bars represent Standard Error of the Mean (SEM). * p

    Journal: PLoS ONE

    Article Title: Protection of the Transplant Kidney from Preservation Injury by Inhibition of Matrix Metalloproteinases

    doi: 10.1371/journal.pone.0157508

    Figure Lengend Snippet: Effect of doxycycline on LDH, cytochrome c oxidase, and total protein release. A: Confirmation of inhibition of MMP-2 and MMP-9 activity by doxycycline using zymography. B: LDH activity in perfusate of rat kidney with warm ischemic injury at 5, 10, and 22 hours perfused with KPS-1 with and without doxycycline (n = 7 each). C: Cytochrome c oxidase level in perfusate of rat kidney with warm ischemic injury at 5, 10, and 22 hours perfused with KPS-1 with and without doxycycline (n = 7 each). D: Total protein release in perfusate of rat kidney with warm ischemic injury at 5, 10, and 22 hours perfused with KPS-1 with and without doxycycline (n = 6 each). Error bars represent Standard Error of the Mean (SEM). * p

    Article Snippet: Inhibition of MMP-2 with siRNA A stock solution of a mixture containing a pool of 3 target-specific 19–25 nucleotide small interfering RNAs designed to knock down rat MMP-2 gene expression (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used (10 μM concentration in a buffer containing 10 μM Tris-HCl, 20 mM NaCl, and 1 mM EDTA at pH 8.0).

    Techniques: Inhibition, Activity Assay, Zymography

    Comparison of MMP-9 and MMP-2 levels at 5, 10, and 22 hours in perfusates from rat kidney with warm ischemic injury undergoing machine cold perfusion (n = 7). Insert: representative zymography with MMP-9 and MMP-2 activitiesError bars represent Standard Error of the Mean (SEM). * p

    Journal: PLoS ONE

    Article Title: Protection of the Transplant Kidney from Preservation Injury by Inhibition of Matrix Metalloproteinases

    doi: 10.1371/journal.pone.0157508

    Figure Lengend Snippet: Comparison of MMP-9 and MMP-2 levels at 5, 10, and 22 hours in perfusates from rat kidney with warm ischemic injury undergoing machine cold perfusion (n = 7). Insert: representative zymography with MMP-9 and MMP-2 activitiesError bars represent Standard Error of the Mean (SEM). * p

    Article Snippet: Inhibition of MMP-2 with siRNA A stock solution of a mixture containing a pool of 3 target-specific 19–25 nucleotide small interfering RNAs designed to knock down rat MMP-2 gene expression (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used (10 μM concentration in a buffer containing 10 μM Tris-HCl, 20 mM NaCl, and 1 mM EDTA at pH 8.0).

    Techniques: Zymography

    NGAL in perfusate of rat kidney with warm ischemic injury at 22 hours with KPS-1 and KPS-1 supplemented with doxycycline and MMP-2 siRNA compared to control (5 hours) (n = 4). Error bars represent Standard Error of the Mean (SEM). * p

    Journal: PLoS ONE

    Article Title: Protection of the Transplant Kidney from Preservation Injury by Inhibition of Matrix Metalloproteinases

    doi: 10.1371/journal.pone.0157508

    Figure Lengend Snippet: NGAL in perfusate of rat kidney with warm ischemic injury at 22 hours with KPS-1 and KPS-1 supplemented with doxycycline and MMP-2 siRNA compared to control (5 hours) (n = 4). Error bars represent Standard Error of the Mean (SEM). * p

    Article Snippet: Inhibition of MMP-2 with siRNA A stock solution of a mixture containing a pool of 3 target-specific 19–25 nucleotide small interfering RNAs designed to knock down rat MMP-2 gene expression (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used (10 μM concentration in a buffer containing 10 μM Tris-HCl, 20 mM NaCl, and 1 mM EDTA at pH 8.0).

    Techniques:

    VEGF promotes increased MMP-2 activity by increasing MT1-MMP expression in ECs. A : MT1-MMP and TIMP-2 mRNA levels, normalized to β-actin mRNA after exposure of iHUVECs to hypoxia for 0–8 h, reported as fold induction compared with untreated controls. B : MT1-MMP and TIMP-2 mRNA levels, normalized to β-actin mRNA after treatment of iHUVECs with increasing doses of rhVEGF for 6 h or increasing duration of treatment with 10 ng rhVEGF, reported as fold induction compared with untreated controls. C : Western blot for MT1-MMP in iHUVECs treated with increasing doses of rhVEGF (for 8 h) or increasing duration of treatment (with 10 ng rhVEGF). GAPDH was used as a loading control. D : MT1-MMP and TIMP-2 mRNA levels (RT-PCR) in iHUVECs treated with conditioned media from MIO-M1 cells exposed to hypoxia (0–6 h) with or without RNAi against VEGF mRNA reported as fold induction compared with untreated controls. E : MMP-2 enzymatic (gelatinase) activity after treatment of iHUVECs with increasing doses of rhVEGF (for 6 h) or increasing duration of treatment (with 10 ng rhVEGF), reported as fold induction compared with untreated controls. All experiments were performed in duplicate and are representative of at least three independent experiments. n = 3 animals in each group. Student t test: * P

    Journal: Diabetes

    Article Title: VEGF Secreted by Hypoxic Müller Cells Induces MMP-2 Expression and Activity in Endothelial Cells to Promote Retinal Neovascularization in Proliferative Diabetic Retinopathy

    doi: 10.2337/db13-0014

    Figure Lengend Snippet: VEGF promotes increased MMP-2 activity by increasing MT1-MMP expression in ECs. A : MT1-MMP and TIMP-2 mRNA levels, normalized to β-actin mRNA after exposure of iHUVECs to hypoxia for 0–8 h, reported as fold induction compared with untreated controls. B : MT1-MMP and TIMP-2 mRNA levels, normalized to β-actin mRNA after treatment of iHUVECs with increasing doses of rhVEGF for 6 h or increasing duration of treatment with 10 ng rhVEGF, reported as fold induction compared with untreated controls. C : Western blot for MT1-MMP in iHUVECs treated with increasing doses of rhVEGF (for 8 h) or increasing duration of treatment (with 10 ng rhVEGF). GAPDH was used as a loading control. D : MT1-MMP and TIMP-2 mRNA levels (RT-PCR) in iHUVECs treated with conditioned media from MIO-M1 cells exposed to hypoxia (0–6 h) with or without RNAi against VEGF mRNA reported as fold induction compared with untreated controls. E : MMP-2 enzymatic (gelatinase) activity after treatment of iHUVECs with increasing doses of rhVEGF (for 6 h) or increasing duration of treatment (with 10 ng rhVEGF), reported as fold induction compared with untreated controls. All experiments were performed in duplicate and are representative of at least three independent experiments. n = 3 animals in each group. Student t test: * P

    Article Snippet: Immunohistochemical detection of HIF-1α (Abcam), MMP-2 (Santa Cruz Biotechnology), CD31 (BD Pharmingen; BD Biosciences), and CD34 (Covance) was performed in paraffin-embedded human tissue and cryopreserved mouse tissue sections using the ABC system (Dako) as previously described ( ).

    Techniques: Activity Assay, Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction

    VEGF is necessary and sufficient for Müller cells to induce MMP-2 mRNA expression in ECs. A : Representative immunofluorescence analysis of VEGF in the retina of P13 OIR eyes compared with P13 control eyes. The EC marker CD31 highlights retinal vasculature. B : Vegf mRNA levels by RT-PCR in primary murine Müller cells exposed to hypoxia (normalized to cyclophilin B mRNA) are reported as fold induction compared with untreated controls. C : VEGF mRNA and protein secretion in MIO-M1 cells exposed to hypoxia reported as fold induction compared with untreated controls. D : VEGF and MMP-2 mRNA levels in iHUVECs treated with conditioned media from MIO-M1 cells exposed to hypoxia (0–6 h) with or without RNAi against VEGF mRNA reported as fold induction compared with untreated controls. E and F : MMP-2 mRNA ( E ) and secreted protein ( F ) levels after treatment of iHUVECs with increasing doses of rhVEGF (for 6 h) or increasing duration of treatment (with 10 ng rhVEGF) reported as fold induction compared with untreated controls. G : MMP-9 mRNA levels after treatment of iHUVECs with increasing doses of rhVEGF (for 6 h) or increasing duration of treatment (with 10 ng rhVEGF) reported as fold induction compared with untreated controls. All experiments were performed in duplicate and are representative of at least three independent experiments. n = 3 animals in each group. Student t test: * P

    Journal: Diabetes

    Article Title: VEGF Secreted by Hypoxic Müller Cells Induces MMP-2 Expression and Activity in Endothelial Cells to Promote Retinal Neovascularization in Proliferative Diabetic Retinopathy

    doi: 10.2337/db13-0014

    Figure Lengend Snippet: VEGF is necessary and sufficient for Müller cells to induce MMP-2 mRNA expression in ECs. A : Representative immunofluorescence analysis of VEGF in the retina of P13 OIR eyes compared with P13 control eyes. The EC marker CD31 highlights retinal vasculature. B : Vegf mRNA levels by RT-PCR in primary murine Müller cells exposed to hypoxia (normalized to cyclophilin B mRNA) are reported as fold induction compared with untreated controls. C : VEGF mRNA and protein secretion in MIO-M1 cells exposed to hypoxia reported as fold induction compared with untreated controls. D : VEGF and MMP-2 mRNA levels in iHUVECs treated with conditioned media from MIO-M1 cells exposed to hypoxia (0–6 h) with or without RNAi against VEGF mRNA reported as fold induction compared with untreated controls. E and F : MMP-2 mRNA ( E ) and secreted protein ( F ) levels after treatment of iHUVECs with increasing doses of rhVEGF (for 6 h) or increasing duration of treatment (with 10 ng rhVEGF) reported as fold induction compared with untreated controls. G : MMP-9 mRNA levels after treatment of iHUVECs with increasing doses of rhVEGF (for 6 h) or increasing duration of treatment (with 10 ng rhVEGF) reported as fold induction compared with untreated controls. All experiments were performed in duplicate and are representative of at least three independent experiments. n = 3 animals in each group. Student t test: * P

    Article Snippet: Immunohistochemical detection of HIF-1α (Abcam), MMP-2 (Santa Cruz Biotechnology), CD31 (BD Pharmingen; BD Biosciences), and CD34 (Covance) was performed in paraffin-embedded human tissue and cryopreserved mouse tissue sections using the ABC system (Dako) as previously described ( ).

    Techniques: Expressing, Immunofluorescence, Marker, Reverse Transcription Polymerase Chain Reaction

    Stabilization of HIF-1α in hypoxic Müller cells does not lead to MMP-2 expression. A : Representative immunofluorescence analysis of HIF-1α in the retina of P13 OIR eyes compared with control P13 eyes. B : Western blot and immunofluorescence for HIF-1α in primary murine Müller cells exposed to hypoxia. GAPDH was used as a loading control. C : Mmp-2 mRNA levels from primary murine Müller cells exposed to hypoxia, normalized to cyclophilin B mRNA, and reported as fold induction compared with cells exposed to 20% O 2 . D and E : Similar studies were performed using MIO-M1 cells. All experiments were performed in duplicate are representative of at least three independent experiments. n = 3 animals in each group. The differences in the fold induction in C and E were not statistically significant ( P > 0.05).

    Journal: Diabetes

    Article Title: VEGF Secreted by Hypoxic Müller Cells Induces MMP-2 Expression and Activity in Endothelial Cells to Promote Retinal Neovascularization in Proliferative Diabetic Retinopathy

    doi: 10.2337/db13-0014

    Figure Lengend Snippet: Stabilization of HIF-1α in hypoxic Müller cells does not lead to MMP-2 expression. A : Representative immunofluorescence analysis of HIF-1α in the retina of P13 OIR eyes compared with control P13 eyes. B : Western blot and immunofluorescence for HIF-1α in primary murine Müller cells exposed to hypoxia. GAPDH was used as a loading control. C : Mmp-2 mRNA levels from primary murine Müller cells exposed to hypoxia, normalized to cyclophilin B mRNA, and reported as fold induction compared with cells exposed to 20% O 2 . D and E : Similar studies were performed using MIO-M1 cells. All experiments were performed in duplicate are representative of at least three independent experiments. n = 3 animals in each group. The differences in the fold induction in C and E were not statistically significant ( P > 0.05).

    Article Snippet: Immunohistochemical detection of HIF-1α (Abcam), MMP-2 (Santa Cruz Biotechnology), CD31 (BD Pharmingen; BD Biosciences), and CD34 (Covance) was performed in paraffin-embedded human tissue and cryopreserved mouse tissue sections using the ABC system (Dako) as previously described ( ).

    Techniques: Expressing, Immunofluorescence, Western Blot

    Increased MMP-2 expression in patients with PDR. A : Representative images from immunohistochemical analysis of MMP-2 expression in NV (black arrows) but not in normal retinal vessels (blue arrow) in the retina of four of four eyes with NV and a known diagnosis of PDR (see inset). No primary antibody was used for the negative control. B : VEGF levels in the aqueous humor of nondiabetic patients, diabetic patients without DR, and diabetic patients with PDR. C : MMP-2 levels in the aqueous humor of these patients. D : MMP-2 levels in the aqueous humor of nondiabetic patients compared with diabetic patients with PDR who were not previously treated with PRP or those who had previous PRP treatment. E , left : VEGF levels in the serum of nondiabetic patients and diabetic patients with PDR. E , right : MMP-2 levels in the serum of nondiabetic patients, diabetic patients without DR, and diabetic patients with PDR. All experiments were performed in duplicate. Control, nondiabetic patients; Diabetic, diabetic patients without DR; PDR, diabetic patients with PDR; Untreated, diabetic patients with PDR who were not previously treated with PRP; Treated, diabetic patients who had received previous PRP treatment. Student t test: * P

    Journal: Diabetes

    Article Title: VEGF Secreted by Hypoxic Müller Cells Induces MMP-2 Expression and Activity in Endothelial Cells to Promote Retinal Neovascularization in Proliferative Diabetic Retinopathy

    doi: 10.2337/db13-0014

    Figure Lengend Snippet: Increased MMP-2 expression in patients with PDR. A : Representative images from immunohistochemical analysis of MMP-2 expression in NV (black arrows) but not in normal retinal vessels (blue arrow) in the retina of four of four eyes with NV and a known diagnosis of PDR (see inset). No primary antibody was used for the negative control. B : VEGF levels in the aqueous humor of nondiabetic patients, diabetic patients without DR, and diabetic patients with PDR. C : MMP-2 levels in the aqueous humor of these patients. D : MMP-2 levels in the aqueous humor of nondiabetic patients compared with diabetic patients with PDR who were not previously treated with PRP or those who had previous PRP treatment. E , left : VEGF levels in the serum of nondiabetic patients and diabetic patients with PDR. E , right : MMP-2 levels in the serum of nondiabetic patients, diabetic patients without DR, and diabetic patients with PDR. All experiments were performed in duplicate. Control, nondiabetic patients; Diabetic, diabetic patients without DR; PDR, diabetic patients with PDR; Untreated, diabetic patients with PDR who were not previously treated with PRP; Treated, diabetic patients who had received previous PRP treatment. Student t test: * P

    Article Snippet: Immunohistochemical detection of HIF-1α (Abcam), MMP-2 (Santa Cruz Biotechnology), CD31 (BD Pharmingen; BD Biosciences), and CD34 (Covance) was performed in paraffin-embedded human tissue and cryopreserved mouse tissue sections using the ABC system (Dako) as previously described ( ).

    Techniques: Expressing, Immunohistochemistry, Negative Control

    HIF-dependent MMP-2 expression during the ischemic phase in the OIR model. A : RT-PCR of Mmp-2 and Mmp-9 mRNA from the neurosensory retina of OIR animals at P12–P15 normalized to cyclophilin B mRNA and reported as fold induction compared with P12. B : Representative Western blot of MMP-2 protein accumulation from the neurosensory retina of OIR animals at P12–P15. Paired lanes represents lysates from two eyes from two separate animals. GAPDH was used as a loading control. C : Representative immunohistochemical analysis of MMP-2 (black arrows; blue chromogenic substrate) in the retina of OIR eyes during the ischemic phase at P14 (posterior retina, left ; peripheral retina, middle ) and during the neovascular phase at P17 ( right ). D : Representative immunohistochemical analysis of HIF-1α (brown chromogenic substrate) in the retina of P13 OIR eyes ( middle ) compared with P13 control eyes ( left ). HIF-1α expression was inhibited with daily intraperitoneal injections of digoxin ( right ). E : RT-PCR of Mmp-2 , Mmp-9 , and Vegf mRNA from the neurosensory retina of OIR animals at P12–P14 with daily intraperitoneal injection of digoxin (+dig) or without injection, normalized to cyclophilin B mRNA, and reported as fold induction compared with P12. All experiments were performed in duplicate and are representative of at least three independent experiments. n = 3 animals in each group. Student t test: * P

    Journal: Diabetes

    Article Title: VEGF Secreted by Hypoxic Müller Cells Induces MMP-2 Expression and Activity in Endothelial Cells to Promote Retinal Neovascularization in Proliferative Diabetic Retinopathy

    doi: 10.2337/db13-0014

    Figure Lengend Snippet: HIF-dependent MMP-2 expression during the ischemic phase in the OIR model. A : RT-PCR of Mmp-2 and Mmp-9 mRNA from the neurosensory retina of OIR animals at P12–P15 normalized to cyclophilin B mRNA and reported as fold induction compared with P12. B : Representative Western blot of MMP-2 protein accumulation from the neurosensory retina of OIR animals at P12–P15. Paired lanes represents lysates from two eyes from two separate animals. GAPDH was used as a loading control. C : Representative immunohistochemical analysis of MMP-2 (black arrows; blue chromogenic substrate) in the retina of OIR eyes during the ischemic phase at P14 (posterior retina, left ; peripheral retina, middle ) and during the neovascular phase at P17 ( right ). D : Representative immunohistochemical analysis of HIF-1α (brown chromogenic substrate) in the retina of P13 OIR eyes ( middle ) compared with P13 control eyes ( left ). HIF-1α expression was inhibited with daily intraperitoneal injections of digoxin ( right ). E : RT-PCR of Mmp-2 , Mmp-9 , and Vegf mRNA from the neurosensory retina of OIR animals at P12–P14 with daily intraperitoneal injection of digoxin (+dig) or without injection, normalized to cyclophilin B mRNA, and reported as fold induction compared with P12. All experiments were performed in duplicate and are representative of at least three independent experiments. n = 3 animals in each group. Student t test: * P

    Article Snippet: Immunohistochemical detection of HIF-1α (Abcam), MMP-2 (Santa Cruz Biotechnology), CD31 (BD Pharmingen; BD Biosciences), and CD34 (Covance) was performed in paraffin-embedded human tissue and cryopreserved mouse tissue sections using the ABC system (Dako) as previously described ( ).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Immunohistochemistry, Injection

    Stabilization of HIF-1α in hypoxic vascular ECs leads to increased MMP-2 mRNA levels. A : Representative immunofluorescence analysis of MMP-2 in the retina of P13 and P14 OIR eyes compared with P13 control eyes. Retinal vasculature was labeled with anti-CD31, an EC marker. B : Western blot and immunofluorescence for HIF-1α in iHUVECs exposed to hypoxia. GAPDH was used as a loading control. C : VEGF and MMP-2 mRNA normalized to β-actin mRNA from iHUVECs exposed to hypoxia, reported as fold induction compared with control cells exposed to 20% O 2 . D , left : Western blot for HIF-1α ( top ) and VEGF mRNA normalized to β-actin mRNA and reported as fold induction compared with control cells exposed to 20% O 2 ( bottom ) from iHUVECs exposed to hypoxia that had been pretreated with digoxin. GAPDH was used as a loading control for the Western blot. D , right : MMP-2 mRNA normalized to β-actin mRNA from iHUVECs exposed to hypoxia, pretreated with digoxin, and reported as fold induction compared with control cells. E , left : Western blot for HIF-1α ( top ) and VEGF mRNA normalized to β-actin mRNA and reported as fold induction compared with control cells ( bottom ) in iHUVECs treated with 1,4-DPCA and DMOG or DFO for 8 h. GAPDH was used as a loading control for the Western blot. E , right : MMP-2 mRNA normalized to β-actin mRNA from iHUVECs treated with 1,4-DPCA, DMOG, or DFO for 8 h, and reported as fold induction compared with control cells. All experiments were performed in duplicate are representative of at least three independent experiments. n = 3 animals in each group. Student t test: * P

    Journal: Diabetes

    Article Title: VEGF Secreted by Hypoxic Müller Cells Induces MMP-2 Expression and Activity in Endothelial Cells to Promote Retinal Neovascularization in Proliferative Diabetic Retinopathy

    doi: 10.2337/db13-0014

    Figure Lengend Snippet: Stabilization of HIF-1α in hypoxic vascular ECs leads to increased MMP-2 mRNA levels. A : Representative immunofluorescence analysis of MMP-2 in the retina of P13 and P14 OIR eyes compared with P13 control eyes. Retinal vasculature was labeled with anti-CD31, an EC marker. B : Western blot and immunofluorescence for HIF-1α in iHUVECs exposed to hypoxia. GAPDH was used as a loading control. C : VEGF and MMP-2 mRNA normalized to β-actin mRNA from iHUVECs exposed to hypoxia, reported as fold induction compared with control cells exposed to 20% O 2 . D , left : Western blot for HIF-1α ( top ) and VEGF mRNA normalized to β-actin mRNA and reported as fold induction compared with control cells exposed to 20% O 2 ( bottom ) from iHUVECs exposed to hypoxia that had been pretreated with digoxin. GAPDH was used as a loading control for the Western blot. D , right : MMP-2 mRNA normalized to β-actin mRNA from iHUVECs exposed to hypoxia, pretreated with digoxin, and reported as fold induction compared with control cells. E , left : Western blot for HIF-1α ( top ) and VEGF mRNA normalized to β-actin mRNA and reported as fold induction compared with control cells ( bottom ) in iHUVECs treated with 1,4-DPCA and DMOG or DFO for 8 h. GAPDH was used as a loading control for the Western blot. E , right : MMP-2 mRNA normalized to β-actin mRNA from iHUVECs treated with 1,4-DPCA, DMOG, or DFO for 8 h, and reported as fold induction compared with control cells. All experiments were performed in duplicate are representative of at least three independent experiments. n = 3 animals in each group. Student t test: * P

    Article Snippet: Immunohistochemical detection of HIF-1α (Abcam), MMP-2 (Santa Cruz Biotechnology), CD31 (BD Pharmingen; BD Biosciences), and CD34 (Covance) was performed in paraffin-embedded human tissue and cryopreserved mouse tissue sections using the ABC system (Dako) as previously described ( ).

    Techniques: Immunofluorescence, Labeling, Marker, Western Blot

    Cooperation between retinal Müller glial cells and ECs increases MMP-2 mRNA levels in cocultured cells. Representative immunofluorescent staining for the EC marker CD31 and the glial cell marker GFAP in the retina of adult ( A ) and OIR ( B ) mice. C : MMP-2 mRNA levels by RT-PCR in cocultures of MIO-M1 cells with iHUVECs under normoxic or hypoxic conditions. D : iHUVECs were cultured at 20% O 2 in the presence of conditioned media (for 4 or 8 h) from MIO-M1 cells exposed to 20% O 2 or 1% O 2 (for 12–48 h, as indicated). Levels of MMP-2 mRNA (normalized to β-actin mRNA) are reported as fold induction compared with untreated controls. All experiments were performed in duplicate and are representative of at least three independent experiments. n = 3 animals in each group. Student t test: * P

    Journal: Diabetes

    Article Title: VEGF Secreted by Hypoxic Müller Cells Induces MMP-2 Expression and Activity in Endothelial Cells to Promote Retinal Neovascularization in Proliferative Diabetic Retinopathy

    doi: 10.2337/db13-0014

    Figure Lengend Snippet: Cooperation between retinal Müller glial cells and ECs increases MMP-2 mRNA levels in cocultured cells. Representative immunofluorescent staining for the EC marker CD31 and the glial cell marker GFAP in the retina of adult ( A ) and OIR ( B ) mice. C : MMP-2 mRNA levels by RT-PCR in cocultures of MIO-M1 cells with iHUVECs under normoxic or hypoxic conditions. D : iHUVECs were cultured at 20% O 2 in the presence of conditioned media (for 4 or 8 h) from MIO-M1 cells exposed to 20% O 2 or 1% O 2 (for 12–48 h, as indicated). Levels of MMP-2 mRNA (normalized to β-actin mRNA) are reported as fold induction compared with untreated controls. All experiments were performed in duplicate and are representative of at least three independent experiments. n = 3 animals in each group. Student t test: * P

    Article Snippet: Immunohistochemical detection of HIF-1α (Abcam), MMP-2 (Santa Cruz Biotechnology), CD31 (BD Pharmingen; BD Biosciences), and CD34 (Covance) was performed in paraffin-embedded human tissue and cryopreserved mouse tissue sections using the ABC system (Dako) as previously described ( ).

    Techniques: Staining, Marker, Mouse Assay, Reverse Transcription Polymerase Chain Reaction, Cell Culture

    VEGF promotes increased MMP-2 expression in retinal NV in vivo and in human PDR tissue. A , top : Schematic demonstrating intravitreal injections of rmVEGF or PBS control. A , bottom : Mmp-2 mRNA from the neurosensory retina of animals injected with rmVEGF over time normalized to cyclophilin B mRNA and reported as fold induction compared with control (PBS-injected) eyes. Representative immunofluorescence analysis of MMP-2 (green arrows) in cross-section ( B ) and flat mount ( C ) from eyes injected with rmVEGF or PBS (control). The EC marker CD31 highlights tip cells of retinal microvasculature (arrowheads). All experiments were performed in duplicate and are representative of at least three independent experiments. n = 3 animals in each group. Student t test: * P

    Journal: Diabetes

    Article Title: VEGF Secreted by Hypoxic Müller Cells Induces MMP-2 Expression and Activity in Endothelial Cells to Promote Retinal Neovascularization in Proliferative Diabetic Retinopathy

    doi: 10.2337/db13-0014

    Figure Lengend Snippet: VEGF promotes increased MMP-2 expression in retinal NV in vivo and in human PDR tissue. A , top : Schematic demonstrating intravitreal injections of rmVEGF or PBS control. A , bottom : Mmp-2 mRNA from the neurosensory retina of animals injected with rmVEGF over time normalized to cyclophilin B mRNA and reported as fold induction compared with control (PBS-injected) eyes. Representative immunofluorescence analysis of MMP-2 (green arrows) in cross-section ( B ) and flat mount ( C ) from eyes injected with rmVEGF or PBS (control). The EC marker CD31 highlights tip cells of retinal microvasculature (arrowheads). All experiments were performed in duplicate and are representative of at least three independent experiments. n = 3 animals in each group. Student t test: * P

    Article Snippet: Immunohistochemical detection of HIF-1α (Abcam), MMP-2 (Santa Cruz Biotechnology), CD31 (BD Pharmingen; BD Biosciences), and CD34 (Covance) was performed in paraffin-embedded human tissue and cryopreserved mouse tissue sections using the ABC system (Dako) as previously described ( ).

    Techniques: Expressing, In Vivo, Injection, Immunofluorescence, Marker

    p-MMP-2 combined with radiation enhances apoptosis in vivo . A, Immunohistochemical analysis of brain sections using anti-MMP-2, anti-VEGF and anti-pFAK antibodies. Sections were photographed (60×). Also shown is the negative control where the primary antibody was replaced by non-specific IgG (insets). B, Tissue sections of mice were evaluated with the TUNEL assay according to manufacturer's instructions and photographed under fluorescent microscopy (60×). For the negative control, samples were incubated with label solution (without terminal transferase) instead of TUNEL reaction mixture (insets). C, siRNA against MMP-2 inhibits U251 tumor cell invasion in vivo . H E staining was performed according to standard protocol, and representative pictures of tumor sections from mock, pSV, p-MMP-2-treated mice are shown (20× and 60×). D , Immunohistochemical analysis of brain sections using anti-human nuclei (HuNu) antibody, a histological marker for identification of human cells (a specific human nuclear antigen). Entire brain sections were photographed (4×; middle row); shown on the top row is a non-tumor region (40×; top row); and shown on the bottom row is tumor and non-tumor overlapping region (40×; bottom row). Also shown is the negative control where the primary antibody was replaced by non-specific IgG (inset).

    Journal: PLoS ONE

    Article Title: MMP-2 siRNA Inhibits Radiation-Enhanced Invasiveness in Glioma Cells

    doi: 10.1371/journal.pone.0020614

    Figure Lengend Snippet: p-MMP-2 combined with radiation enhances apoptosis in vivo . A, Immunohistochemical analysis of brain sections using anti-MMP-2, anti-VEGF and anti-pFAK antibodies. Sections were photographed (60×). Also shown is the negative control where the primary antibody was replaced by non-specific IgG (insets). B, Tissue sections of mice were evaluated with the TUNEL assay according to manufacturer's instructions and photographed under fluorescent microscopy (60×). For the negative control, samples were incubated with label solution (without terminal transferase) instead of TUNEL reaction mixture (insets). C, siRNA against MMP-2 inhibits U251 tumor cell invasion in vivo . H E staining was performed according to standard protocol, and representative pictures of tumor sections from mock, pSV, p-MMP-2-treated mice are shown (20× and 60×). D , Immunohistochemical analysis of brain sections using anti-human nuclei (HuNu) antibody, a histological marker for identification of human cells (a specific human nuclear antigen). Entire brain sections were photographed (4×; middle row); shown on the top row is a non-tumor region (40×; top row); and shown on the bottom row is tumor and non-tumor overlapping region (40×; bottom row). Also shown is the negative control where the primary antibody was replaced by non-specific IgG (inset).

    Article Snippet: Then, slides were incubated with 0.1% Triton X-100, blocked with 3% BSA in PBS, and incubated with anti-MMP-2, anti-VEGF, anti-pFAK (Santa Cruz Biotechnology, SantaCruz, CA) or anti-human nuclei (HuNu) antibody (Millipore, Temecula, CA) (1∶10 dilution).

    Techniques: In Vivo, Immunohistochemistry, Negative Control, Mouse Assay, TUNEL Assay, Microscopy, Incubation, Staining, Marker

    p-MMP-2 inhibits colony formation and induces apoptosis and activation of pro-apoptotic molecules in glioma cells. A, Clonogenic assay was performed as described in Materials and Methods . The cells were cultured, and colonies larger than 50 cells were counted. Columns : mean of triplicate experiments; bars : SD; * p

    Journal: PLoS ONE

    Article Title: MMP-2 siRNA Inhibits Radiation-Enhanced Invasiveness in Glioma Cells

    doi: 10.1371/journal.pone.0020614

    Figure Lengend Snippet: p-MMP-2 inhibits colony formation and induces apoptosis and activation of pro-apoptotic molecules in glioma cells. A, Clonogenic assay was performed as described in Materials and Methods . The cells were cultured, and colonies larger than 50 cells were counted. Columns : mean of triplicate experiments; bars : SD; * p

    Article Snippet: Then, slides were incubated with 0.1% Triton X-100, blocked with 3% BSA in PBS, and incubated with anti-MMP-2, anti-VEGF, anti-pFAK (Santa Cruz Biotechnology, SantaCruz, CA) or anti-human nuclei (HuNu) antibody (Millipore, Temecula, CA) (1∶10 dilution).

    Techniques: Activation Assay, Clonogenic Assay, Cell Culture

    p-MMP-2 transfection inhibits radiation-enhanced glioma cell invasion. U-251 and U-87 cells were transfected with mock, p-SV or p-MMP-2, and irradiated as described earlier. Cells were trypsinized and counted, and 5×10 5 cells from each treatment condition were allowed to invade transwell inserts containing 12-µm-pore polycarbonate membranes pre-coated with Matrigel for 24 h at 37°C. Afterwards, cells were fixed and stained with Hema-3. Cells that had migrated to the lower side of the membrane were photographed under a light microscope at 20× magnification. Percentages of invading cells were quantified by counting five fields from each treatment condition. Columns : mean of triplicate experiments; bars : SD; * p

    Journal: PLoS ONE

    Article Title: MMP-2 siRNA Inhibits Radiation-Enhanced Invasiveness in Glioma Cells

    doi: 10.1371/journal.pone.0020614

    Figure Lengend Snippet: p-MMP-2 transfection inhibits radiation-enhanced glioma cell invasion. U-251 and U-87 cells were transfected with mock, p-SV or p-MMP-2, and irradiated as described earlier. Cells were trypsinized and counted, and 5×10 5 cells from each treatment condition were allowed to invade transwell inserts containing 12-µm-pore polycarbonate membranes pre-coated with Matrigel for 24 h at 37°C. Afterwards, cells were fixed and stained with Hema-3. Cells that had migrated to the lower side of the membrane were photographed under a light microscope at 20× magnification. Percentages of invading cells were quantified by counting five fields from each treatment condition. Columns : mean of triplicate experiments; bars : SD; * p

    Article Snippet: Then, slides were incubated with 0.1% Triton X-100, blocked with 3% BSA in PBS, and incubated with anti-MMP-2, anti-VEGF, anti-pFAK (Santa Cruz Biotechnology, SantaCruz, CA) or anti-human nuclei (HuNu) antibody (Millipore, Temecula, CA) (1∶10 dilution).

    Techniques: Transfection, Irradiation, Staining, Light Microscopy

    p-MMP-2 transfection in combination with radiation inhibits glioma cell migration. U-251 and U-87 cells were cultured for formation of spheroids as described in Materials and Methods . Spheroids were then transfected with mock, p-SV or p-MMP-2, and followed by irradiation as described earlier. At the end of the migration assay, spheroids were fixed and stained with Hema-3. Migration of cells from spheroids to monolayers was measured using a microscope calibrated with a stage and ocular micrometer. Columns : mean of triplicate experiments; bars : SD; * p

    Journal: PLoS ONE

    Article Title: MMP-2 siRNA Inhibits Radiation-Enhanced Invasiveness in Glioma Cells

    doi: 10.1371/journal.pone.0020614

    Figure Lengend Snippet: p-MMP-2 transfection in combination with radiation inhibits glioma cell migration. U-251 and U-87 cells were cultured for formation of spheroids as described in Materials and Methods . Spheroids were then transfected with mock, p-SV or p-MMP-2, and followed by irradiation as described earlier. At the end of the migration assay, spheroids were fixed and stained with Hema-3. Migration of cells from spheroids to monolayers was measured using a microscope calibrated with a stage and ocular micrometer. Columns : mean of triplicate experiments; bars : SD; * p

    Article Snippet: Then, slides were incubated with 0.1% Triton X-100, blocked with 3% BSA in PBS, and incubated with anti-MMP-2, anti-VEGF, anti-pFAK (Santa Cruz Biotechnology, SantaCruz, CA) or anti-human nuclei (HuNu) antibody (Millipore, Temecula, CA) (1∶10 dilution).

    Techniques: Transfection, Migration, Cell Culture, Irradiation, Staining, Microscopy

    p-MMP-2 combined with radiation inhibits tumor growth in vivo . U-251 (1×10 6 ) cells were injected intracerebrally into athymic mice. After ten days, animals were separated into five groups and were treated on alternate days with intracerebral injections of p-SV or p-MMP-2 for a total of 4 doses (60 µg per dose) and 2 doses of radiation (4 Gy per dose) as described in Materials and Methods . Six weeks after the experiment was initiated, mice were euthanized with intracardiac perfusion of PBS, followed by formaldehyde. The brains were then removed. A, Six weeks after the experiment was initiated, an intraperitonal injection of 2.5 mg D-luciferin sodium salt diluted in 50 µL of PBS was given, and animals were photographed under the IVIS camera for fluorescent light emission. The brains were removed and fixed in 10% phosphate-buffered formaldehyde, and the fixed tissue samples were then processed into paraffin blocks. Brain sections (5 µM thick) were stained with hematoxylin and eosin (H E), and photographed under a light microscope (4× and 40×). B, Every fifth or sixth brain section (5 µM thick) was stained with H E solution, and the tumor masses (H E-stained) were manually traced on the microscope attached computer screen. Areas were calculated using Image Pro Discovery Program software (Media Cybernetics, Inc., Silver Spring, MD). The total tumor volume was calculated as the summed area on all slices, multiplied by the slice separation. Columns : mean of area of tumor portion of all mice in the group (n = 8); bars : SD; * p

    Journal: PLoS ONE

    Article Title: MMP-2 siRNA Inhibits Radiation-Enhanced Invasiveness in Glioma Cells

    doi: 10.1371/journal.pone.0020614

    Figure Lengend Snippet: p-MMP-2 combined with radiation inhibits tumor growth in vivo . U-251 (1×10 6 ) cells were injected intracerebrally into athymic mice. After ten days, animals were separated into five groups and were treated on alternate days with intracerebral injections of p-SV or p-MMP-2 for a total of 4 doses (60 µg per dose) and 2 doses of radiation (4 Gy per dose) as described in Materials and Methods . Six weeks after the experiment was initiated, mice were euthanized with intracardiac perfusion of PBS, followed by formaldehyde. The brains were then removed. A, Six weeks after the experiment was initiated, an intraperitonal injection of 2.5 mg D-luciferin sodium salt diluted in 50 µL of PBS was given, and animals were photographed under the IVIS camera for fluorescent light emission. The brains were removed and fixed in 10% phosphate-buffered formaldehyde, and the fixed tissue samples were then processed into paraffin blocks. Brain sections (5 µM thick) were stained with hematoxylin and eosin (H E), and photographed under a light microscope (4× and 40×). B, Every fifth or sixth brain section (5 µM thick) was stained with H E solution, and the tumor masses (H E-stained) were manually traced on the microscope attached computer screen. Areas were calculated using Image Pro Discovery Program software (Media Cybernetics, Inc., Silver Spring, MD). The total tumor volume was calculated as the summed area on all slices, multiplied by the slice separation. Columns : mean of area of tumor portion of all mice in the group (n = 8); bars : SD; * p

    Article Snippet: Then, slides were incubated with 0.1% Triton X-100, blocked with 3% BSA in PBS, and incubated with anti-MMP-2, anti-VEGF, anti-pFAK (Santa Cruz Biotechnology, SantaCruz, CA) or anti-human nuclei (HuNu) antibody (Millipore, Temecula, CA) (1∶10 dilution).

    Techniques: In Vivo, Injection, Mouse Assay, Staining, Light Microscopy, Microscopy, Software

    Radiation enhances MMP-2 and p-MMP-2 inhibits MMP-2 activity and expression in glioma cell lines. A, U-251 and U-87 cells were irradiated with 0–12 Gy X-ray, incubated for 24 h, and conditioned medium collected. MMP-2 activity was determined by gelatin zymography. The band intensities of MMP-2 activity were quantified by densitometry. Columns : mean of triplicate experiments; bars : SD; * p

    Journal: PLoS ONE

    Article Title: MMP-2 siRNA Inhibits Radiation-Enhanced Invasiveness in Glioma Cells

    doi: 10.1371/journal.pone.0020614

    Figure Lengend Snippet: Radiation enhances MMP-2 and p-MMP-2 inhibits MMP-2 activity and expression in glioma cell lines. A, U-251 and U-87 cells were irradiated with 0–12 Gy X-ray, incubated for 24 h, and conditioned medium collected. MMP-2 activity was determined by gelatin zymography. The band intensities of MMP-2 activity were quantified by densitometry. Columns : mean of triplicate experiments; bars : SD; * p

    Article Snippet: Then, slides were incubated with 0.1% Triton X-100, blocked with 3% BSA in PBS, and incubated with anti-MMP-2, anti-VEGF, anti-pFAK (Santa Cruz Biotechnology, SantaCruz, CA) or anti-human nuclei (HuNu) antibody (Millipore, Temecula, CA) (1∶10 dilution).

    Techniques: Activity Assay, Expressing, Irradiation, Incubation, Zymography

    p-MMP-2 transfection inhibits radiation-enhanced MMP-2 activity and expression levels as well as cell viability. A, U-251 and U-87 cells were transfected with mock (PBS), p-SV or p-MMP-2 (2 µg), and after 72 h of incubation, cells were irradiated with 0, 2, 4, 6 or 8 Gy and incubated for a further 24 h. Conditioned media was used to determine MMP-2 activity by gelatin zymography, and total cell lysates were used to determine MMP-2 levels by Western blotting. B, Total RNA was used to determine MMP-2 mRNA transcription levels by RT-PCR with gene-specific primers. GAPDH served as a loading control. C, U-251 and U-87 cells were transfected with mock, p-SV or p-MMP-2 and irradiated as described above. 24 h after radiation, the cells were fixed and processed to visualize MMP-2 expression. The cells were mounted using mounting media with DAPI to visualize the nucleus. D, U-251 and U-87 cells were transfected with mock, p-SV or p-MMP-2, and irradiated for 72 h after transfection. After a another 24 h of incubation, cell viability was analyzed by MTT assay (absorbance read at 550 nm). Columns : mean of triplicate experiments; bars : SD; * p

    Journal: PLoS ONE

    Article Title: MMP-2 siRNA Inhibits Radiation-Enhanced Invasiveness in Glioma Cells

    doi: 10.1371/journal.pone.0020614

    Figure Lengend Snippet: p-MMP-2 transfection inhibits radiation-enhanced MMP-2 activity and expression levels as well as cell viability. A, U-251 and U-87 cells were transfected with mock (PBS), p-SV or p-MMP-2 (2 µg), and after 72 h of incubation, cells were irradiated with 0, 2, 4, 6 or 8 Gy and incubated for a further 24 h. Conditioned media was used to determine MMP-2 activity by gelatin zymography, and total cell lysates were used to determine MMP-2 levels by Western blotting. B, Total RNA was used to determine MMP-2 mRNA transcription levels by RT-PCR with gene-specific primers. GAPDH served as a loading control. C, U-251 and U-87 cells were transfected with mock, p-SV or p-MMP-2 and irradiated as described above. 24 h after radiation, the cells were fixed and processed to visualize MMP-2 expression. The cells were mounted using mounting media with DAPI to visualize the nucleus. D, U-251 and U-87 cells were transfected with mock, p-SV or p-MMP-2, and irradiated for 72 h after transfection. After a another 24 h of incubation, cell viability was analyzed by MTT assay (absorbance read at 550 nm). Columns : mean of triplicate experiments; bars : SD; * p

    Article Snippet: Then, slides were incubated with 0.1% Triton X-100, blocked with 3% BSA in PBS, and incubated with anti-MMP-2, anti-VEGF, anti-pFAK (Santa Cruz Biotechnology, SantaCruz, CA) or anti-human nuclei (HuNu) antibody (Millipore, Temecula, CA) (1∶10 dilution).

    Techniques: Transfection, Activity Assay, Expressing, Incubation, Irradiation, Zymography, Western Blot, Reverse Transcription Polymerase Chain Reaction, MTT Assay

    p-MMP-2 inhibits radiation-enhanced tumor culture medium-induced microtubule network formation in endothelial cells and downregulates expression of angiogenesis-associated molecules. A, Human microvascular endothelial cells (5×10 4 ) were seeded in 96-well plates and cultured with conditioned medium collected from U-251 and U-87 glioma cells transfected with mock, p-SV, and p-MMP-2, and irradiated as described earlier. 24 h after radiation treatment, the cells were washed, fixed and stained with Hema-3 and photographed. Percentages of branches were quantified by counting five fields in each condition. Columns : mean of triplicate experiments; bars : SD; * p

    Journal: PLoS ONE

    Article Title: MMP-2 siRNA Inhibits Radiation-Enhanced Invasiveness in Glioma Cells

    doi: 10.1371/journal.pone.0020614

    Figure Lengend Snippet: p-MMP-2 inhibits radiation-enhanced tumor culture medium-induced microtubule network formation in endothelial cells and downregulates expression of angiogenesis-associated molecules. A, Human microvascular endothelial cells (5×10 4 ) were seeded in 96-well plates and cultured with conditioned medium collected from U-251 and U-87 glioma cells transfected with mock, p-SV, and p-MMP-2, and irradiated as described earlier. 24 h after radiation treatment, the cells were washed, fixed and stained with Hema-3 and photographed. Percentages of branches were quantified by counting five fields in each condition. Columns : mean of triplicate experiments; bars : SD; * p

    Article Snippet: Then, slides were incubated with 0.1% Triton X-100, blocked with 3% BSA in PBS, and incubated with anti-MMP-2, anti-VEGF, anti-pFAK (Santa Cruz Biotechnology, SantaCruz, CA) or anti-human nuclei (HuNu) antibody (Millipore, Temecula, CA) (1∶10 dilution).

    Techniques: Expressing, Cell Culture, Transfection, Irradiation, Staining