vascular endothelial growth factor vegf  (Thermo Fisher)


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    Vascular Endothelial Growth Factor VEGF Ab 7
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    ms-1467-pcl
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    Structured Review

    Thermo Fisher vascular endothelial growth factor vegf
    (a) EMA immunostain highlighting whorls of meningioma cells (200x); (b) intense and diffuse <t>VEGF</t> expression in an anaplastic meningioma (200x); (c) <t>Ki67</t> immunostain revealing a moderate proliferative activity in a case of atypical meningioma (200x); (d) benign meningioma with significant PR expression (200x); (e) CD20 highlighting rare B lymphocytes (100x); (f) CD3 showing a large population of T lymphocytes (100x).

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    Images

    1) Product Images from "Biological and Demographic Profile of Meningiomas in a Cohort of Egyptian Patients: Impact on Tumor Recurrence"

    Article Title: Biological and Demographic Profile of Meningiomas in a Cohort of Egyptian Patients: Impact on Tumor Recurrence

    Journal: The Scientific World Journal

    doi: 10.1155/2013/375139

    (a) EMA immunostain highlighting whorls of meningioma cells (200x); (b) intense and diffuse VEGF expression in an anaplastic meningioma (200x); (c) Ki67 immunostain revealing a moderate proliferative activity in a case of atypical meningioma (200x); (d) benign meningioma with significant PR expression (200x); (e) CD20 highlighting rare B lymphocytes (100x); (f) CD3 showing a large population of T lymphocytes (100x).
    Figure Legend Snippet: (a) EMA immunostain highlighting whorls of meningioma cells (200x); (b) intense and diffuse VEGF expression in an anaplastic meningioma (200x); (c) Ki67 immunostain revealing a moderate proliferative activity in a case of atypical meningioma (200x); (d) benign meningioma with significant PR expression (200x); (e) CD20 highlighting rare B lymphocytes (100x); (f) CD3 showing a large population of T lymphocytes (100x).

    Techniques Used: Expressing, Activity Assay

    2) Product Images from "Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress"

    Article Title: Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress

    Journal: Stem Cells International

    doi: 10.1155/2018/9682856

    Early passage ASCs overexpress the neurotropic protein HGF but not VEGF nor the proinflammatory cytokine IL-1 β . ASCs at passage 3 that were cultured in serum-free conditions for 48 hours were compared to the control group of ASCs at passage 5. Both cells and medium were collected and analyzed at mRNA level and at protein level by qRT-PCR and by ELISA, respectively. (a) qRT-PCR analysis of HGF, VEGF, and IL-1 β . (b) ELISA for HGF protein levels. Each experiment was performed a minimum of 3 samples from 3 different patients. Each experiment was performed a minimum of 3 times.
    Figure Legend Snippet: Early passage ASCs overexpress the neurotropic protein HGF but not VEGF nor the proinflammatory cytokine IL-1 β . ASCs at passage 3 that were cultured in serum-free conditions for 48 hours were compared to the control group of ASCs at passage 5. Both cells and medium were collected and analyzed at mRNA level and at protein level by qRT-PCR and by ELISA, respectively. (a) qRT-PCR analysis of HGF, VEGF, and IL-1 β . (b) ELISA for HGF protein levels. Each experiment was performed a minimum of 3 samples from 3 different patients. Each experiment was performed a minimum of 3 times.

    Techniques Used: Cell Culture, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    3) Product Images from "Exosome Delivered Anticancer Drugs Across the Blood-Brain Barrier for Brain Cancer Therapy in Danio Rerio"

    Article Title: Exosome Delivered Anticancer Drugs Across the Blood-Brain Barrier for Brain Cancer Therapy in Danio Rerio

    Journal: Pharmaceutical research

    doi: 10.1007/s11095-014-1593-y

    Inhibitory of VEGF in vivo using bEND.3 exosome delivered doxorubicin on zebrafish cancer model (* Results are significantly different, p
    Figure Legend Snippet: Inhibitory of VEGF in vivo using bEND.3 exosome delivered doxorubicin on zebrafish cancer model (* Results are significantly different, p

    Techniques Used: In Vivo

    4) Product Images from "Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress"

    Article Title: Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress

    Journal: Stem Cells International

    doi: 10.1155/2018/9682856

    Early passage ASCs overexpress the neurotropic protein HGF but not VEGF nor the proinflammatory cytokine IL-1 β . ASCs at passage 3 that were cultured in serum-free conditions for 48 hours were compared to the control group of ASCs at passage 5. Both cells and medium were collected and analyzed at mRNA level and at protein level by qRT-PCR and by ELISA, respectively. (a) qRT-PCR analysis of HGF, VEGF, and IL-1 β . (b) ELISA for HGF protein levels. Each experiment was performed a minimum of 3 samples from 3 different patients. Each experiment was performed a minimum of 3 times.
    Figure Legend Snippet: Early passage ASCs overexpress the neurotropic protein HGF but not VEGF nor the proinflammatory cytokine IL-1 β . ASCs at passage 3 that were cultured in serum-free conditions for 48 hours were compared to the control group of ASCs at passage 5. Both cells and medium were collected and analyzed at mRNA level and at protein level by qRT-PCR and by ELISA, respectively. (a) qRT-PCR analysis of HGF, VEGF, and IL-1 β . (b) ELISA for HGF protein levels. Each experiment was performed a minimum of 3 samples from 3 different patients. Each experiment was performed a minimum of 3 times.

    Techniques Used: Cell Culture, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    5) Product Images from "Conditioned media from endothelial progenitor cells cultured in simulated microgravity promote angiogenesis and bone fracture healing"

    Article Title: Conditioned media from endothelial progenitor cells cultured in simulated microgravity promote angiogenesis and bone fracture healing

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-020-02074-y

    MG-CM facilitated neovascularization of fracture areas in vivo. a Microfil perfusion was performed to examine the newly formed vessels of the regenerated areas followed by micro-CT scan. b Total vessel volume was calculated and analyzed based on reconstructive vessel images. c – h Immunohistochemistry staining was employed to detect the expression of CD31, VEGF-A, and MMP-9 in regenerative areas followed by quantitative analyses. Abbreviations: NG-CM, conditioned media from endothelial progenitor cells under normal gravity; MG-CM, conditioned media from endothelial progenitor cells under microgravity; VEGF-A, vascular endothelial growth factor A; MMP-9, matrix metalloproteinase 9. Data were presented as the mean ± standard deviation. * P
    Figure Legend Snippet: MG-CM facilitated neovascularization of fracture areas in vivo. a Microfil perfusion was performed to examine the newly formed vessels of the regenerated areas followed by micro-CT scan. b Total vessel volume was calculated and analyzed based on reconstructive vessel images. c – h Immunohistochemistry staining was employed to detect the expression of CD31, VEGF-A, and MMP-9 in regenerative areas followed by quantitative analyses. Abbreviations: NG-CM, conditioned media from endothelial progenitor cells under normal gravity; MG-CM, conditioned media from endothelial progenitor cells under microgravity; VEGF-A, vascular endothelial growth factor A; MMP-9, matrix metalloproteinase 9. Data were presented as the mean ± standard deviation. * P

    Techniques Used: In Vivo, Micro-CT, Immunohistochemistry, Staining, Expressing, Standard Deviation

    MG suppressed EPCs’ proliferation but activated HIF-1α/eNOS/NO axis. a MTT assay was performed to detect EPCs’ proliferation under MG or NG exposure. b Immunofluorescence staining showed the expression levels of Ki67 under NG (left) or MG (right) exposure. DAPI was for nuclear counterstain. c A cluster of angiogenic genes expression was examined by qRT-PCR after 12-, 24- and 48-h exposures of NG and MG, and GAPDH was used as the internal reference. d Western blot was employed to identify the expression of HIF-1α, eNOS and iNOS at protein levels. e Griess assay was performed to detect the NO contents in MG-CM and NG-CM after 12-, 24-, and 48-h exposures, and the concentrations were normalized to cell numbers. This experiment was repeated independently three times. Abbreviations: EPCs, endothelial progenitor cells; MG, microgravity; NG, normal gravity; DAPI, 4, 6-diamidino-2-phenylindole; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HIF-1α, hypoxia-induced factor-1α; eNOS, endothelial nitric oxide synthase; iNOS, inducible nitric oxide synthase; NO, nitric oxide; VEGF, vascular endothelial growth factor; MMP-9, matrix metalloproteinase 9; PDGF-B, platelet-derived growth factor-B; Ang-2, angiogenin-2. Data were presented as the mean ± standard deviation. * P
    Figure Legend Snippet: MG suppressed EPCs’ proliferation but activated HIF-1α/eNOS/NO axis. a MTT assay was performed to detect EPCs’ proliferation under MG or NG exposure. b Immunofluorescence staining showed the expression levels of Ki67 under NG (left) or MG (right) exposure. DAPI was for nuclear counterstain. c A cluster of angiogenic genes expression was examined by qRT-PCR after 12-, 24- and 48-h exposures of NG and MG, and GAPDH was used as the internal reference. d Western blot was employed to identify the expression of HIF-1α, eNOS and iNOS at protein levels. e Griess assay was performed to detect the NO contents in MG-CM and NG-CM after 12-, 24-, and 48-h exposures, and the concentrations were normalized to cell numbers. This experiment was repeated independently three times. Abbreviations: EPCs, endothelial progenitor cells; MG, microgravity; NG, normal gravity; DAPI, 4, 6-diamidino-2-phenylindole; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HIF-1α, hypoxia-induced factor-1α; eNOS, endothelial nitric oxide synthase; iNOS, inducible nitric oxide synthase; NO, nitric oxide; VEGF, vascular endothelial growth factor; MMP-9, matrix metalloproteinase 9; PDGF-B, platelet-derived growth factor-B; Ang-2, angiogenin-2. Data were presented as the mean ± standard deviation. * P

    Techniques Used: MTT Assay, Immunofluorescence, Staining, Expressing, Quantitative RT-PCR, Western Blot, Griess Assay, Derivative Assay, Standard Deviation

    NO-induced activation of FAK/Erk1/2-MAPK signaling pathway contributed to enhanced pro-angiogenic properties of MG-CM. a Western blot was employed to detect the phosphorylation levels of FAK and Erk1/2. b The phosphorylation level of Erk1/2 was examined after Erk1/2-MAPK-selective inhibitor PD98059 application for 4 h. c , d Tube formation assay was performed to detect the angiogenic abilities of HUVECs when PD98059 was added or not, followed by quantitative analysis. e Immunofluorescence staining results showed the expression of Ki67 and CD31 in different groups. DAPI was for nuclear counterstain. f The mRNA expression level of VEGF was detected by qRT-PCR, and GAPDH was used as internal reference. g The schematic illustration of MG-induced HIF-1α/eNOS/NO activation of EPCs promoting HUVECs’ proliferation, migration, and angiogenesis. This experiment was repeated independently three times. Abbreviations: EPCs, endothelial progenitor cells; HUVECs, human umbilical vein endothelial cells; FAK, focal adhesion kinase; NG-CM, conditioned media from endothelial progenitor cells under normal gravity; MG-CM, conditioned media from endothelial progenitor cells under microgravity; NO, nitric oxide; DAPI, 4, 6-diamidino-2-phenylindole; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; VEGF, vascular endothelial growth factor; HIF-1α, hypoxia-induced factor-1α; eNOS, endothelial nitric oxide synthase. Data were presented as the mean ± standard deviation. * P
    Figure Legend Snippet: NO-induced activation of FAK/Erk1/2-MAPK signaling pathway contributed to enhanced pro-angiogenic properties of MG-CM. a Western blot was employed to detect the phosphorylation levels of FAK and Erk1/2. b The phosphorylation level of Erk1/2 was examined after Erk1/2-MAPK-selective inhibitor PD98059 application for 4 h. c , d Tube formation assay was performed to detect the angiogenic abilities of HUVECs when PD98059 was added or not, followed by quantitative analysis. e Immunofluorescence staining results showed the expression of Ki67 and CD31 in different groups. DAPI was for nuclear counterstain. f The mRNA expression level of VEGF was detected by qRT-PCR, and GAPDH was used as internal reference. g The schematic illustration of MG-induced HIF-1α/eNOS/NO activation of EPCs promoting HUVECs’ proliferation, migration, and angiogenesis. This experiment was repeated independently three times. Abbreviations: EPCs, endothelial progenitor cells; HUVECs, human umbilical vein endothelial cells; FAK, focal adhesion kinase; NG-CM, conditioned media from endothelial progenitor cells under normal gravity; MG-CM, conditioned media from endothelial progenitor cells under microgravity; NO, nitric oxide; DAPI, 4, 6-diamidino-2-phenylindole; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; VEGF, vascular endothelial growth factor; HIF-1α, hypoxia-induced factor-1α; eNOS, endothelial nitric oxide synthase. Data were presented as the mean ± standard deviation. * P

    Techniques Used: Activation Assay, Western Blot, Tube Formation Assay, Immunofluorescence, Staining, Expressing, Quantitative RT-PCR, Migration, Standard Deviation

    MG-CM promoted HUVECs’ proliferation, migration, and angiogenesis in vitro partially through increased NO production. a The effects of DMEM, NG-CM, MG-CM, and NO reduced MG-CM on HUVECs’ proliferation were analyzed by MTT assay. b The Ki67 expression in HUVECs was detected by immunofluorescence staining. DAPI was for nuclear counterstain. c – f Transwell assay and wound healing assay were employed to detect the migration capacity changes of HUVECs after treatment followed by quantitative analyses. g The CD31 expression levels of HUVECs in each group were detected by immunofluorescence staining. h , i Tube formation assay was used to detect the angiogenic abilities of HUVECs followed by quantitative analysis. j qRT-PCR results showed the relative expression levels of VEGF and MMP-9, with GAPDH as the internal reference. This experiment was repeated independently three times. Abbreviations: HUVECs, human umbilical vein endothelial cells; NG-CM, conditioned media from endothelial progenitor cells under normal gravity; MG-CM, conditioned media from endothelial progenitor cells under microgravity; NO, nitric oxide; DAPI, 4, 6-diamidino-2-phenylindole; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; VEGF, vascular endothelial growth factor; MMP-9, matrix metalloproteinase 9. Data were presented as the mean ± standard deviation. * P
    Figure Legend Snippet: MG-CM promoted HUVECs’ proliferation, migration, and angiogenesis in vitro partially through increased NO production. a The effects of DMEM, NG-CM, MG-CM, and NO reduced MG-CM on HUVECs’ proliferation were analyzed by MTT assay. b The Ki67 expression in HUVECs was detected by immunofluorescence staining. DAPI was for nuclear counterstain. c – f Transwell assay and wound healing assay were employed to detect the migration capacity changes of HUVECs after treatment followed by quantitative analyses. g The CD31 expression levels of HUVECs in each group were detected by immunofluorescence staining. h , i Tube formation assay was used to detect the angiogenic abilities of HUVECs followed by quantitative analysis. j qRT-PCR results showed the relative expression levels of VEGF and MMP-9, with GAPDH as the internal reference. This experiment was repeated independently three times. Abbreviations: HUVECs, human umbilical vein endothelial cells; NG-CM, conditioned media from endothelial progenitor cells under normal gravity; MG-CM, conditioned media from endothelial progenitor cells under microgravity; NO, nitric oxide; DAPI, 4, 6-diamidino-2-phenylindole; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; VEGF, vascular endothelial growth factor; MMP-9, matrix metalloproteinase 9. Data were presented as the mean ± standard deviation. * P

    Techniques Used: Migration, In Vitro, MTT Assay, Expressing, Immunofluorescence, Staining, Transwell Assay, Wound Healing Assay, Tube Formation Assay, Quantitative RT-PCR, Standard Deviation

    6) Product Images from "Xanthohumol Impairs Human Prostate Cancer Cell Growth and Invasion and Diminishes the Incidence and Progression of Advanced Tumors in TRAMP Mice"

    Article Title: Xanthohumol Impairs Human Prostate Cancer Cell Growth and Invasion and Diminishes the Incidence and Progression of Advanced Tumors in TRAMP Mice

    Journal: Molecular Medicine

    doi: 10.2119/molmed.2012.00174

    XN inhibits FAK, AKT and NF-κB activation and controls VEGF release and the migratory potential of PCa cells. Western blot analyses show a remarkable decrease of constitutive FAK phosphorylation after 5 h of treatment with XN (A), which correlates
    Figure Legend Snippet: XN inhibits FAK, AKT and NF-κB activation and controls VEGF release and the migratory potential of PCa cells. Western blot analyses show a remarkable decrease of constitutive FAK phosphorylation after 5 h of treatment with XN (A), which correlates

    Techniques Used: Activation Assay, Western Blot

    7) Product Images from "Inducible Costimulator (ICOS) and ICOS Ligand Signaling Has Pivotal Roles in Skin Wound Healing via Cytokine Production"

    Article Title: Inducible Costimulator (ICOS) and ICOS Ligand Signaling Has Pivotal Roles in Skin Wound Healing via Cytokine Production

    Journal: The American Journal of Pathology

    doi: 10.1016/j.ajpath.2011.07.048

    Cytokines, chemokines, and growth factors in wounds from wild-type (WT) and mutant mice. A: Expression of IL-4, IL-6, IL-10, TNF-α, CTGF, PDGF, TGF-β, and VEGF mRNAs measured by quantitative RT-PCR. B: Concentrations of IL-6, IL-10, TNF-α,
    Figure Legend Snippet: Cytokines, chemokines, and growth factors in wounds from wild-type (WT) and mutant mice. A: Expression of IL-4, IL-6, IL-10, TNF-α, CTGF, PDGF, TGF-β, and VEGF mRNAs measured by quantitative RT-PCR. B: Concentrations of IL-6, IL-10, TNF-α,

    Techniques Used: Mutagenesis, Mouse Assay, Expressing, Quantitative RT-PCR

    8) Product Images from "CYR61, a product of a growth factor-inducible immediate early gene, promotes angiogenesis and tumor growth"

    Article Title: CYR61, a product of a growth factor-inducible immediate early gene, promotes angiogenesis and tumor growth

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi:

    Chemotactic activity of CYR61. The migration of HMVECs was measured in a modified Boyden chamber assay. The cells placed in the lower chamber that migrated into the upper chamber were counted in three high power fields for each condition after a 4-hr incubation at 37°C. CYR61 was used at 1 μg/ml (except for dose response in A ), vitronectin was used at 5 μg/ml, bFGF was used at 10 ng/ml, and VEGF was used at 1 ng/ml as chemoattractants. ( A ) CYR61-stimulated cell migration is dose-dependent. Cells that migrated into the upper well, where the indicated amount of purified CYR61 (▪) or the equivalent amount of CYR61 storage buffer (○) was placed, were counted. In parallel, HMVEC migration was measured by using bFGF as a chemoattractant, placed in the upper well with various amounts of CYR61 storage buffer corresponding to the amounts used in CYR61-stimulated migration experiment. Cells migrated are expressed as a percentage of bFGF-induced migration ± SD ( B ) Specific inhibition of CYR61-stimulated cell migration by anti-Cyr61 antibodies (25 μg/ml). HMVEC migration was monitored as described above by using either CYR61 or bFGF as a chemoattractant. Where indicated, these proteins were preincubated with anti-Cyr61 antibodies before use. ( C ) Directed migration of HMVEC tested in a checkerboard-type analysis. CYR61 or bFGF was added to the upper chamber, the lower chamber, neither chamber, or both chambers as indicated. In B and C , results are shown as the number of cells that migrated per three high power fields ± SD ( D ) Specific inhibition of CYR61-stimulated migration by an antibody against integrin α V β 3 but not α V β 5 . HMVEC migration was monitored by using CYR61, vitronectin, bFGF, or VEGF as chemoattractants. Where indicated, cells were preincubated with 50 μg/ml LM609 for 1 hr before addition to the lower chamber. Results are expressed as percentage of cells migrated to VEGF. Neg., background migration in the absence of chemoattractant.
    Figure Legend Snippet: Chemotactic activity of CYR61. The migration of HMVECs was measured in a modified Boyden chamber assay. The cells placed in the lower chamber that migrated into the upper chamber were counted in three high power fields for each condition after a 4-hr incubation at 37°C. CYR61 was used at 1 μg/ml (except for dose response in A ), vitronectin was used at 5 μg/ml, bFGF was used at 10 ng/ml, and VEGF was used at 1 ng/ml as chemoattractants. ( A ) CYR61-stimulated cell migration is dose-dependent. Cells that migrated into the upper well, where the indicated amount of purified CYR61 (▪) or the equivalent amount of CYR61 storage buffer (○) was placed, were counted. In parallel, HMVEC migration was measured by using bFGF as a chemoattractant, placed in the upper well with various amounts of CYR61 storage buffer corresponding to the amounts used in CYR61-stimulated migration experiment. Cells migrated are expressed as a percentage of bFGF-induced migration ± SD ( B ) Specific inhibition of CYR61-stimulated cell migration by anti-Cyr61 antibodies (25 μg/ml). HMVEC migration was monitored as described above by using either CYR61 or bFGF as a chemoattractant. Where indicated, these proteins were preincubated with anti-Cyr61 antibodies before use. ( C ) Directed migration of HMVEC tested in a checkerboard-type analysis. CYR61 or bFGF was added to the upper chamber, the lower chamber, neither chamber, or both chambers as indicated. In B and C , results are shown as the number of cells that migrated per three high power fields ± SD ( D ) Specific inhibition of CYR61-stimulated migration by an antibody against integrin α V β 3 but not α V β 5 . HMVEC migration was monitored by using CYR61, vitronectin, bFGF, or VEGF as chemoattractants. Where indicated, cells were preincubated with 50 μg/ml LM609 for 1 hr before addition to the lower chamber. Results are expressed as percentage of cells migrated to VEGF. Neg., background migration in the absence of chemoattractant.

    Techniques Used: Activity Assay, Migration, Modification, Boyden Chamber Assay, Incubation, Purification, Inhibition

    9) Product Images from "Nano-biological mesh constructed by astragaloside-IV-induced bone marrow mesenchymal stem cells on PLGA-NPs-SIS can be used for abdominal wall reconstruction"

    Article Title: Nano-biological mesh constructed by astragaloside-IV-induced bone marrow mesenchymal stem cells on PLGA-NPs-SIS can be used for abdominal wall reconstruction

    Journal: American Journal of Translational Research

    doi:

    Protein and mRNA expression of inflammatory and angiogenic factors. A. qRT-PCR analysis of COL1A1, COL3A1, VEGF, MMP-9, MCP-1, and IL-6 levels; relative mRNA levels are based on GAPDH expression. B. WB analysis of the expression of indicated proteins (GAPDH was adopted as a loading control). *P
    Figure Legend Snippet: Protein and mRNA expression of inflammatory and angiogenic factors. A. qRT-PCR analysis of COL1A1, COL3A1, VEGF, MMP-9, MCP-1, and IL-6 levels; relative mRNA levels are based on GAPDH expression. B. WB analysis of the expression of indicated proteins (GAPDH was adopted as a loading control). *P

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot

    10) Product Images from "Metabolic Syndrome Triggered by Fructose Diet Impairs Neuronal Function and Vascular Integrity in ApoE-KO Mouse Retinas: Implications of Autophagy Deficient Activation"

    Article Title: Metabolic Syndrome Triggered by Fructose Diet Impairs Neuronal Function and Vascular Integrity in ApoE-KO Mouse Retinas: Implications of Autophagy Deficient Activation

    Journal: Frontiers in Cell and Developmental Biology

    doi: 10.3389/fcell.2020.573987

    Vascular analysis. (A) Representative photomicrographs (scale of 100 μm, 100x magnification) of GSA-IB4 labeled (green) blood vessels in flat-mounts of the central area of whole retinas in the different experimental groups at 4 months of diet (n = 3). (B) Scheme of quantification (top left panel), bars represent the average vascular density (% area, bottom left panel), the average vascular diameter (μm, top right panel) and the average vascular branching (bottom right panel) of flat-mounted whole retinas of mice from the different experimental groups at 4 months of diet. (C) representative photomicrographs (scale 100 μm, 50x magnification) of α-SMA labeled (red) blood vessels in flat-mounts of the central area of whole retinas of mice from the different experimental groups at 4 months of diet ( n = 4). Bars represent the average vascular density (% area). (D) Bars represent the average VEGF (top panel) and HIF-1α (bottom panel) mRNA levels relative to actin in mouse retinal extracts of the different experimental groups at 4 months of diet ( n = 4). (E) Representative photomicrographs (scale 500 μm, 100x magnification) of albumin–Evans blue-complex leakage (red) in flat-mounts of the whole retinas in mice from the different experimental groups at 4 months of diet ( n = 3). Square indicate the place of 1000x magnification, scale 250 μm. (F) Bars represent albumin protein relative to actin levels in retinal extracts of mice from the different experimental groups at 4 months of diet (bottom panel, n = 3). Representative blot is showed (top panel). (G) representative photomicrographs (scale of 100 μm, 50x magnification) of GFAP labeled (red) in flat-mounted of central area of the whole retinas of mice from the different experimental groups at 4 months of diet ( n = 3). Squares indicate the place of 500x magnification, scale 500 μm. (H) Bars represent average protein expression of GFAP ( n = 4) and GS ( n = 5) relative to tubulin in retinal extracts of mice from the different experimental groups at 4 months of diet (bottom panel). Representative blot is showed (top panel). (I) Representative photomicrographs (scale 25 μm, 200x magnification) of GFAP label (green) in retinal criosections (10 μm) of mice from the different experimental groups at 4 months of diet ( n = 3). Data correspond to mean ± SEM. Two-way ANOVA followed by Bonferroni post hoc test or Kruskal–Wallis followed by Dunn’s post hoc test. * p
    Figure Legend Snippet: Vascular analysis. (A) Representative photomicrographs (scale of 100 μm, 100x magnification) of GSA-IB4 labeled (green) blood vessels in flat-mounts of the central area of whole retinas in the different experimental groups at 4 months of diet (n = 3). (B) Scheme of quantification (top left panel), bars represent the average vascular density (% area, bottom left panel), the average vascular diameter (μm, top right panel) and the average vascular branching (bottom right panel) of flat-mounted whole retinas of mice from the different experimental groups at 4 months of diet. (C) representative photomicrographs (scale 100 μm, 50x magnification) of α-SMA labeled (red) blood vessels in flat-mounts of the central area of whole retinas of mice from the different experimental groups at 4 months of diet ( n = 4). Bars represent the average vascular density (% area). (D) Bars represent the average VEGF (top panel) and HIF-1α (bottom panel) mRNA levels relative to actin in mouse retinal extracts of the different experimental groups at 4 months of diet ( n = 4). (E) Representative photomicrographs (scale 500 μm, 100x magnification) of albumin–Evans blue-complex leakage (red) in flat-mounts of the whole retinas in mice from the different experimental groups at 4 months of diet ( n = 3). Square indicate the place of 1000x magnification, scale 250 μm. (F) Bars represent albumin protein relative to actin levels in retinal extracts of mice from the different experimental groups at 4 months of diet (bottom panel, n = 3). Representative blot is showed (top panel). (G) representative photomicrographs (scale of 100 μm, 50x magnification) of GFAP labeled (red) in flat-mounted of central area of the whole retinas of mice from the different experimental groups at 4 months of diet ( n = 3). Squares indicate the place of 500x magnification, scale 500 μm. (H) Bars represent average protein expression of GFAP ( n = 4) and GS ( n = 5) relative to tubulin in retinal extracts of mice from the different experimental groups at 4 months of diet (bottom panel). Representative blot is showed (top panel). (I) Representative photomicrographs (scale 25 μm, 200x magnification) of GFAP label (green) in retinal criosections (10 μm) of mice from the different experimental groups at 4 months of diet ( n = 3). Data correspond to mean ± SEM. Two-way ANOVA followed by Bonferroni post hoc test or Kruskal–Wallis followed by Dunn’s post hoc test. * p

    Techniques Used: Labeling, Mouse Assay, Expressing

    11) Product Images from "Effects of amniotic membrane transplantation on cytokines expression in chemically burned rat corneas"

    Article Title: Effects of amniotic membrane transplantation on cytokines expression in chemically burned rat corneas

    Journal: International Journal of Ophthalmology

    doi: 10.3980/j.issn.2222-3959.2011.01.07

    Effects of AMT on the expressions of EGF, bFGF, VEGF and MPP-2 in corneas of rats
    Figure Legend Snippet: Effects of AMT on the expressions of EGF, bFGF, VEGF and MPP-2 in corneas of rats

    Techniques Used:

    12) Product Images from "Candesartan targeting of angiotensin II type 1 receptor demonstrates benefits for hypertension in pregnancy via the NF-κB signaling pathway"

    Article Title: Candesartan targeting of angiotensin II type 1 receptor demonstrates benefits for hypertension in pregnancy via the NF-κB signaling pathway

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2018.9070

    Effects of Candesartan on Ang-IITR and expression levels of hypertension-related proteins in vascular endothelial cells. (A) Angiotensin expression levels in control, healthy or Candesartan-treated vascular endothelial cells. (B) Angiotensin release in vascular endothelial cells after treatment with homocysteine or Candesartan. (C) Ang-IITR expression levels and (D) angiotensin plasma concentration levels in control or Candesartan-treated mice with gestational hypertension. Analysis of (E) VEGF, (F) TGF-β, (G) Ang-1 and (H) PLGF expression levels in control, healthy or Candesartan-treated vascular endothelial cells. The data are presented as the mean ± standard error. *P
    Figure Legend Snippet: Effects of Candesartan on Ang-IITR and expression levels of hypertension-related proteins in vascular endothelial cells. (A) Angiotensin expression levels in control, healthy or Candesartan-treated vascular endothelial cells. (B) Angiotensin release in vascular endothelial cells after treatment with homocysteine or Candesartan. (C) Ang-IITR expression levels and (D) angiotensin plasma concentration levels in control or Candesartan-treated mice with gestational hypertension. Analysis of (E) VEGF, (F) TGF-β, (G) Ang-1 and (H) PLGF expression levels in control, healthy or Candesartan-treated vascular endothelial cells. The data are presented as the mean ± standard error. *P

    Techniques Used: Expressing, Concentration Assay, Mouse Assay

    13) Product Images from "Follistatin‐Like 3 Enhances the Function of Endothelial Cells Derived from Pluripotent Stem Cells by Facilitating β‐Catenin Nuclear Translocation Through Inhibition of Glycogen Synthase Kinase‐3β Activity"

    Article Title: Follistatin‐Like 3 Enhances the Function of Endothelial Cells Derived from Pluripotent Stem Cells by Facilitating β‐Catenin Nuclear Translocation Through Inhibition of Glycogen Synthase Kinase‐3β Activity

    Journal: Stem Cells (Dayton, Ohio)

    doi: 10.1002/stem.2820

    Induced pluripotent stem cell differentiation toward endothelial cells. Human iPS Cells were differentiated using StemPro serum free media supplemented with BMP4, Activin A, fibroblast growth factor (FGF), and vascular endothelial growth factor (VEGF) for 5 days. The differentiated cells were seeded on collagen IV, while CD144 positive cells were magnetically sorted on day 6 using MicroBeads Kit (Miltenyi BIotec) and culturing in EGM‐2 media (LONZA). (A) : Images show morphology of iPS Cells (left panel) and of their differentiated EC counterparts (right panel) Scale bar: 50 μm. (B) : Real time polymerase chain reaction (PCR) data revealed significantly increased early and (C) late EC marker mRNA expression (Data is means ±SEM [ n = 3], *, p
    Figure Legend Snippet: Induced pluripotent stem cell differentiation toward endothelial cells. Human iPS Cells were differentiated using StemPro serum free media supplemented with BMP4, Activin A, fibroblast growth factor (FGF), and vascular endothelial growth factor (VEGF) for 5 days. The differentiated cells were seeded on collagen IV, while CD144 positive cells were magnetically sorted on day 6 using MicroBeads Kit (Miltenyi BIotec) and culturing in EGM‐2 media (LONZA). (A) : Images show morphology of iPS Cells (left panel) and of their differentiated EC counterparts (right panel) Scale bar: 50 μm. (B) : Real time polymerase chain reaction (PCR) data revealed significantly increased early and (C) late EC marker mRNA expression (Data is means ±SEM [ n = 3], *, p

    Techniques Used: Cell Differentiation, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Marker, Expressing

    14) Product Images from "Macrophage Migration Inhibitory Factor Is Involved in Ectopic Endometrial Tissue Growth and Peritoneal-Endometrial Tissue Interaction In Vivo: A Plausible Link to Endometriosis Development"

    Article Title: Macrophage Migration Inhibitory Factor Is Involved in Ectopic Endometrial Tissue Growth and Peritoneal-Endometrial Tissue Interaction In Vivo: A Plausible Link to Endometriosis Development

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0110434

    Real-time PCR analysis of the expression of genes mainly involved in cell adhesion and vascularisation of endometriosis-like lesions. Histogram representation of the effect of MIF genetic depletion or antagonism versus controls on VEGF (A, B), COX2 (C, D), BCL2 (E, F), BAX (G, H), ITGAV (I, J) and ITGB3 (K, L) mRNA expression in endometriosis-like lesions by quantitative real time PCR. For each factor, the ratio of mRNA level to GAPDH mRNA was determined. Results were from WT and KOmice (n = 6) with no treatment (controls) and from WT treated with ISO-1 (n = 5). Data are mean ± SEM; *, p
    Figure Legend Snippet: Real-time PCR analysis of the expression of genes mainly involved in cell adhesion and vascularisation of endometriosis-like lesions. Histogram representation of the effect of MIF genetic depletion or antagonism versus controls on VEGF (A, B), COX2 (C, D), BCL2 (E, F), BAX (G, H), ITGAV (I, J) and ITGB3 (K, L) mRNA expression in endometriosis-like lesions by quantitative real time PCR. For each factor, the ratio of mRNA level to GAPDH mRNA was determined. Results were from WT and KOmice (n = 6) with no treatment (controls) and from WT treated with ISO-1 (n = 5). Data are mean ± SEM; *, p

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing

    15) Product Images from "Influence of the Expression of Inflammatory Markers on Kidney after Fetal Programming in an Experimental Model of Renal Failure"

    Article Title: Influence of the Expression of Inflammatory Markers on Kidney after Fetal Programming in an Experimental Model of Renal Failure

    Journal: Journal of Immunology Research

    doi: 10.1155/2016/9151607

    mRNA gene expression of MCP-1, IL-1, NOS3, TGF- β , TNF- α , and VEGF in the kidney of control offspring and diabetic offspring with or without folic acid, aged two months. (a) The gene expression of MCP-1 was increased in the CFA2 group; (b and c) the gene expression of IL-1 and NOS3 showed no significant difference between the study groups; (d) the gene expression of TGF- β was decreased in the DC2 group; (e and f) the gene expression of TNF- α and VEGF showed no significant difference between the groups. ∗ Significant differences.
    Figure Legend Snippet: mRNA gene expression of MCP-1, IL-1, NOS3, TGF- β , TNF- α , and VEGF in the kidney of control offspring and diabetic offspring with or without folic acid, aged two months. (a) The gene expression of MCP-1 was increased in the CFA2 group; (b and c) the gene expression of IL-1 and NOS3 showed no significant difference between the study groups; (d) the gene expression of TGF- β was decreased in the DC2 group; (e and f) the gene expression of TNF- α and VEGF showed no significant difference between the groups. ∗ Significant differences.

    Techniques Used: Expressing

    mRNA gene expression of MCP-1, IL-1, NOS3, TGF- β , TNF- α , and VEGF in the kidney of control offspring and diabetic offspring with or without folic acid, aged five months. (a, b, c, d, and e) The gene expression of MCP-1, IL-1, NOS3, TGF- β , and TNF- α showed no significant difference between the study groups; (f) the gene expression of VEGF was decreased in the CFA5 and DFA5 groups. ∗ Significant differences.
    Figure Legend Snippet: mRNA gene expression of MCP-1, IL-1, NOS3, TGF- β , TNF- α , and VEGF in the kidney of control offspring and diabetic offspring with or without folic acid, aged five months. (a, b, c, d, and e) The gene expression of MCP-1, IL-1, NOS3, TGF- β , and TNF- α showed no significant difference between the study groups; (f) the gene expression of VEGF was decreased in the CFA5 and DFA5 groups. ∗ Significant differences.

    Techniques Used: Expressing

    16) Product Images from "A clinical and basic study of optimal endometrial preparation protocols for patients with infertility undergoing frozen-thawed embryo transfer"

    Article Title: A clinical and basic study of optimal endometrial preparation protocols for patients with infertility undergoing frozen-thawed embryo transfer

    Journal: Experimental and Therapeutic Medicine

    doi: 10.3892/etm.2020.8914

    Quantitative analysis of mean endometrial LIF, integrin β3, OPN, VEGF, HOXA10and HOXA11mRNA levels of patients who did not undergo embryo transfers on the day of embryo transfer. * P
    Figure Legend Snippet: Quantitative analysis of mean endometrial LIF, integrin β3, OPN, VEGF, HOXA10and HOXA11mRNA levels of patients who did not undergo embryo transfers on the day of embryo transfer. * P

    Techniques Used:

    17) Product Images from "The Sulfamate Small Molecule CAIX Inhibitor S4 Modulates Doxorubicin Efficacy"

    Article Title: The Sulfamate Small Molecule CAIX Inhibitor S4 Modulates Doxorubicin Efficacy

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0161040

    Effect of S4 on doxorubicin efficacy in HT29 –CAIX high and HT29 –CAIX low cells. Exposing HT29 cells to doxycycline (doxy) reduced CA9 mRNA levels during normoxia (N) and hypoxia (H), whereas VEGF levels were unaffected ( A ). CAIX protein levels minimized in HT29 –CAIX low cells when exposed to doxycycline ( B ). Quantification of three independent biological repeats showed minimal residual CAIX protein expression in HT29 –CAIX low cells as compared to parental cells or HT29 –CAIX high cells without a KD ( C ). Cell viability assays of HT29 –CAIX high ( D ) or HT29 –CAIX low cells ( E ) with increasing concentrations of doxorubicin. Cells were exposed to vehicle (black) or S4 (green) during normoxia (N), or to vehicle (red) or S4 (blue) during hypoxia (H). Results of three independent biological repeats are shown (mean ± SEM).
    Figure Legend Snippet: Effect of S4 on doxorubicin efficacy in HT29 –CAIX high and HT29 –CAIX low cells. Exposing HT29 cells to doxycycline (doxy) reduced CA9 mRNA levels during normoxia (N) and hypoxia (H), whereas VEGF levels were unaffected ( A ). CAIX protein levels minimized in HT29 –CAIX low cells when exposed to doxycycline ( B ). Quantification of three independent biological repeats showed minimal residual CAIX protein expression in HT29 –CAIX low cells as compared to parental cells or HT29 –CAIX high cells without a KD ( C ). Cell viability assays of HT29 –CAIX high ( D ) or HT29 –CAIX low cells ( E ) with increasing concentrations of doxorubicin. Cells were exposed to vehicle (black) or S4 (green) during normoxia (N), or to vehicle (red) or S4 (blue) during hypoxia (H). Results of three independent biological repeats are shown (mean ± SEM).

    Techniques Used: Expressing

    18) Product Images from "Role of LAMA4 Gene in Regulating Extravillous Trophoblasts in Pathogenesis of Preeclampsia"

    Article Title: Role of LAMA4 Gene in Regulating Extravillous Trophoblasts in Pathogenesis of Preeclampsia

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    doi: 10.12659/MSM.917402

    VEGF and sFlt-1 mRNA levels. ( A ) RT-PCR band, ( B ) VEGF and sFlt-1 mRNA levels. * p
    Figure Legend Snippet: VEGF and sFlt-1 mRNA levels. ( A ) RT-PCR band, ( B ) VEGF and sFlt-1 mRNA levels. * p

    Techniques Used: Reverse Transcription Polymerase Chain Reaction

    19) Product Images from "Hair growth-promoting effect of Geranium sibiricum extract in human dermal papilla cells and C57BL/6 mice"

    Article Title: Hair growth-promoting effect of Geranium sibiricum extract in human dermal papilla cells and C57BL/6 mice

    Journal: BMC Complementary and Alternative Medicine

    doi: 10.1186/s12906-017-1624-4

    Hair growth-stimulating effect of GSE in an in vivo model. a Observation of C57BL/6 mice hair growth for 3 weeks. b Histological analysis of hair follicles, mast cells, and stem cell factor expression on the back skin of C57BL/6 mice. c The numbers of mast cells were counted by toluidine blue staining. d Expression of vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and transforming growth factor beta 1 (TGF-β1) in the back skin of C57BL/6 mice. The fold changes were normalized to the expression of glyceraldehyde 3-phosphate dehydrogenase; the values are expressed as the mean ± SEM. Values sharing the same superscript letters differ significantly at p
    Figure Legend Snippet: Hair growth-stimulating effect of GSE in an in vivo model. a Observation of C57BL/6 mice hair growth for 3 weeks. b Histological analysis of hair follicles, mast cells, and stem cell factor expression on the back skin of C57BL/6 mice. c The numbers of mast cells were counted by toluidine blue staining. d Expression of vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and transforming growth factor beta 1 (TGF-β1) in the back skin of C57BL/6 mice. The fold changes were normalized to the expression of glyceraldehyde 3-phosphate dehydrogenase; the values are expressed as the mean ± SEM. Values sharing the same superscript letters differ significantly at p

    Techniques Used: In Vivo, Mouse Assay, Expressing, Staining

    Changes in the levels of Ki67 protein, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), and transforming growth factor beta 1 (TGF-β1) in human dermal papilla cells (hDPCs) induced by GSE. a Immunohistochemistry of Ki67. b Numbers of Ki67-positive cells. c Relative expression levels of HGF, VEGF, and TGF-β1 in GSE-treated hDPCs. The fold changes were normalized to the expression of glyceraldehyde 3-phosphate dehydrogenase; the values are expressed as the mean ± SEM. Values sharing the same superscript letters differ significantly at p
    Figure Legend Snippet: Changes in the levels of Ki67 protein, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), and transforming growth factor beta 1 (TGF-β1) in human dermal papilla cells (hDPCs) induced by GSE. a Immunohistochemistry of Ki67. b Numbers of Ki67-positive cells. c Relative expression levels of HGF, VEGF, and TGF-β1 in GSE-treated hDPCs. The fold changes were normalized to the expression of glyceraldehyde 3-phosphate dehydrogenase; the values are expressed as the mean ± SEM. Values sharing the same superscript letters differ significantly at p

    Techniques Used: Immunohistochemistry, Expressing

    20) Product Images from "Topographical cues regulate the crosstalk between MSCs and macrophages"

    Article Title: Topographical cues regulate the crosstalk between MSCs and macrophages

    Journal: Biomaterials

    doi: 10.1016/j.biomaterials.2014.10.028

    Overview of the effect of topographical cues in the soluble factor-guided communication between MSCs and macrophages. MSCs were seeded in 2D substrates (left panel) or 3D scaffolds (right panel) and co-cultured with macrophages using a transwell insert system. The figure illustrates soluble factors whose levels decreased (red), increased (green) or were not affected (blue) in 3D co-cultures as compared to 2D settings. 3D topography sensing activates MSCs to produce PGE 2 and TSG-6 anti-inflammatory proteins while attenuates the secretion of IL-6, MCP-1, RANTES, GM-CSF, M-CSF and RANKL in co-cultures with macrophages. TNF-α, IL-10 and VEGF levels are unaffected by topographical features of the substrate. The lower number of monocytes in 3D settings represents the decrease in monocyte migration to the local inflammatory milieu provided by 3D-arranged MSCs as compared to 2D co-cultures. This attenuation in monocyte recruitment is mediated, at least in part, by reduced levels of IL-6 and MCP-1 (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.).
    Figure Legend Snippet: Overview of the effect of topographical cues in the soluble factor-guided communication between MSCs and macrophages. MSCs were seeded in 2D substrates (left panel) or 3D scaffolds (right panel) and co-cultured with macrophages using a transwell insert system. The figure illustrates soluble factors whose levels decreased (red), increased (green) or were not affected (blue) in 3D co-cultures as compared to 2D settings. 3D topography sensing activates MSCs to produce PGE 2 and TSG-6 anti-inflammatory proteins while attenuates the secretion of IL-6, MCP-1, RANTES, GM-CSF, M-CSF and RANKL in co-cultures with macrophages. TNF-α, IL-10 and VEGF levels are unaffected by topographical features of the substrate. The lower number of monocytes in 3D settings represents the decrease in monocyte migration to the local inflammatory milieu provided by 3D-arranged MSCs as compared to 2D co-cultures. This attenuation in monocyte recruitment is mediated, at least in part, by reduced levels of IL-6 and MCP-1 (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.).

    Techniques Used: Cell Culture, Migration

    21) Product Images from "Intracoronary Administration of Allogeneic Adipose Tissue–Derived Mesenchymal Stem Cells Improves Myocardial Perfusion But Not Left Ventricle Function, in a Translational Model of Acute Myocardial Infarction"

    Article Title: Intracoronary Administration of Allogeneic Adipose Tissue–Derived Mesenchymal Stem Cells Improves Myocardial Perfusion But Not Left Ventricle Function, in a Translational Model of Acute Myocardial Infarction

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    doi: 10.1161/JAHA.117.005771

    Effects of ATMSC s therapy on myocardial vascularization and perfusion. A, Microphotographs on the left (endothelial cells stained in brown and smooth muscle cells stained in red) and bar chart on the right showing increased vessel density in the border infarct zone of animals receiving cell therapy. B, Bar chart on the left and cardiac magnetic resonance images (myocardial quantitative perfusion maps) on the right illustrate an enhancement on myocardial perfusion of infarct border zone in animals receiving allogeneic ATMSC s. Arrows point to perfusion analysis region of interest (gray, anterior infarct border; white, core infarcted area; black, septum infarct border; yellow, posterior remote myocardium). C, Charts show up‐regulation of the pro‐angiogenic mediators SDF ‐1α, GM ‐ CSF , and VEGF in the infarcted tissue of the ATMSC s group compared with vehicle group, as assessed by qPCR . * P
    Figure Legend Snippet: Effects of ATMSC s therapy on myocardial vascularization and perfusion. A, Microphotographs on the left (endothelial cells stained in brown and smooth muscle cells stained in red) and bar chart on the right showing increased vessel density in the border infarct zone of animals receiving cell therapy. B, Bar chart on the left and cardiac magnetic resonance images (myocardial quantitative perfusion maps) on the right illustrate an enhancement on myocardial perfusion of infarct border zone in animals receiving allogeneic ATMSC s. Arrows point to perfusion analysis region of interest (gray, anterior infarct border; white, core infarcted area; black, septum infarct border; yellow, posterior remote myocardium). C, Charts show up‐regulation of the pro‐angiogenic mediators SDF ‐1α, GM ‐ CSF , and VEGF in the infarcted tissue of the ATMSC s group compared with vehicle group, as assessed by qPCR . * P

    Techniques Used: Staining, Real-time Polymerase Chain Reaction

    22) Product Images from "Differences in peritoneal response after exposure to low-GDP bicarbonate/lactate-buffered dialysis solution compared to conventional dialysis solution in a uremic mouse model"

    Article Title: Differences in peritoneal response after exposure to low-GDP bicarbonate/lactate-buffered dialysis solution compared to conventional dialysis solution in a uremic mouse model

    Journal: International Urology and Nephrology

    doi: 10.1007/s11255-018-1872-3

    Exposure to a bicarbonate/lactate-buffered solution is associated with increase of pro-inflammatory cytokines. Protein levels of the main pro-inflammatory cytokines detected in effluents collected from mouse after 8 weeks of PD fluid exposure. Cytokines levels (ρg/ml) are represented as means ± SD ( n ≥ 7) as follows: a TGFβ1, b IL-1β, c IL-6, d TNFα, e VEGF, f INFγ, g IL-17, h IL-4, i IL-5, j MIP-1α, k MIP-1β. C control, LS lactate PD fluid, BLS bicarbonate/lactate PD fluid. Differences were considered statistically significant for P
    Figure Legend Snippet: Exposure to a bicarbonate/lactate-buffered solution is associated with increase of pro-inflammatory cytokines. Protein levels of the main pro-inflammatory cytokines detected in effluents collected from mouse after 8 weeks of PD fluid exposure. Cytokines levels (ρg/ml) are represented as means ± SD ( n ≥ 7) as follows: a TGFβ1, b IL-1β, c IL-6, d TNFα, e VEGF, f INFγ, g IL-17, h IL-4, i IL-5, j MIP-1α, k MIP-1β. C control, LS lactate PD fluid, BLS bicarbonate/lactate PD fluid. Differences were considered statistically significant for P

    Techniques Used:

    23) Product Images from "Differences in peritoneal response after exposure to low-GDP bicarbonate/lactate-buffered dialysis solution compared to conventional dialysis solution in a uremic mouse model"

    Article Title: Differences in peritoneal response after exposure to low-GDP bicarbonate/lactate-buffered dialysis solution compared to conventional dialysis solution in a uremic mouse model

    Journal: International Urology and Nephrology

    doi: 10.1007/s11255-018-1872-3

    Exposure to a bicarbonate/lactate-buffered solution is associated with increase of pro-inflammatory cytokines. Protein levels of the main pro-inflammatory cytokines detected in effluents collected from mouse after 8 weeks of PD fluid exposure. Cytokines levels (ρg/ml) are represented as means ± SD ( n ≥ 7) as follows: a TGFβ1, b IL-1β, c IL-6, d TNFα, e VEGF, f INFγ, g IL-17, h IL-4, i IL-5, j MIP-1α, k MIP-1β. C control, LS lactate PD fluid, BLS bicarbonate/lactate PD fluid. Differences were considered statistically significant for P
    Figure Legend Snippet: Exposure to a bicarbonate/lactate-buffered solution is associated with increase of pro-inflammatory cytokines. Protein levels of the main pro-inflammatory cytokines detected in effluents collected from mouse after 8 weeks of PD fluid exposure. Cytokines levels (ρg/ml) are represented as means ± SD ( n ≥ 7) as follows: a TGFβ1, b IL-1β, c IL-6, d TNFα, e VEGF, f INFγ, g IL-17, h IL-4, i IL-5, j MIP-1α, k MIP-1β. C control, LS lactate PD fluid, BLS bicarbonate/lactate PD fluid. Differences were considered statistically significant for P

    Techniques Used:

    24) Product Images from "Addition of Mesenchymal Stem Cells Enhances the Therapeutic Effects of Skeletal Myoblast Cell-Sheet Transplantation in a Rat Ischemic Cardiomyopathy Model"

    Article Title: Addition of Mesenchymal Stem Cells Enhances the Therapeutic Effects of Skeletal Myoblast Cell-Sheet Transplantation in a Rat Ischemic Cardiomyopathy Model

    Journal: Tissue Engineering. Part A

    doi: 10.1089/ten.tea.2012.0534

    Expression of VEGF and HGF is higher at the infarct area. (A, B) Levels of mRNA in the transplanted infarct and infarct-remote heart areas by real-time PCR using rat-specific primers. The HGF and VEGF mRNA expressions within the transplanted infarct area
    Figure Legend Snippet: Expression of VEGF and HGF is higher at the infarct area. (A, B) Levels of mRNA in the transplanted infarct and infarct-remote heart areas by real-time PCR using rat-specific primers. The HGF and VEGF mRNA expressions within the transplanted infarct area

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    25) Product Images from "Boswellic Acid Inhibits Growth and Metastasis of Human Colorectal Cancer in Orthotopic Mouse Model By Downregulating Inflammatory, Proliferative, Invasive, and Angiogenic Biomarkers"

    Article Title: Boswellic Acid Inhibits Growth and Metastasis of Human Colorectal Cancer in Orthotopic Mouse Model By Downregulating Inflammatory, Proliferative, Invasive, and Angiogenic Biomarkers

    Journal: International Journal of Cancer. Journal International du Cancer

    doi: 10.1002/ijc.26251

    AKBA inhibits the expression of NF-κB and NF-κB-regulated gene products in CRC samples. (A) Electrophoretic mobility shift assay analysis showed the inhibition of NF-κB by AKBA in nuclear extracts from animal tissue. (B) Western blot showing that AKBA inhibited the expression of NF-κB-dependent gene products that regulate proliferation (cyclin D1 and COX-2), invasion (ICAM-1 and MMP-9), metastasis (CXCR4), and angiogenesis (VEGF). AKBA inhibits the expression of antiapoptotic gene products such as IAP-1, Bcl-xL, Bcl-2, and survivin in CRC tissues. Samples from three animals in each group were analyzed, and representative data are shown. V, vehicle; L, low (AKBA 50 mg/kg); M, medium (100 mg/kg); and H, high (200 mg/kg).
    Figure Legend Snippet: AKBA inhibits the expression of NF-κB and NF-κB-regulated gene products in CRC samples. (A) Electrophoretic mobility shift assay analysis showed the inhibition of NF-κB by AKBA in nuclear extracts from animal tissue. (B) Western blot showing that AKBA inhibited the expression of NF-κB-dependent gene products that regulate proliferation (cyclin D1 and COX-2), invasion (ICAM-1 and MMP-9), metastasis (CXCR4), and angiogenesis (VEGF). AKBA inhibits the expression of antiapoptotic gene products such as IAP-1, Bcl-xL, Bcl-2, and survivin in CRC tissues. Samples from three animals in each group were analyzed, and representative data are shown. V, vehicle; L, low (AKBA 50 mg/kg); M, medium (100 mg/kg); and H, high (200 mg/kg).

    Techniques Used: Expressing, Electrophoretic Mobility Shift Assay, Inhibition, Western Blot

    26) Product Images from "Follistatin‐Like 3 Enhances the Function of Endothelial Cells Derived from Pluripotent Stem Cells by Facilitating β‐Catenin Nuclear Translocation Through Inhibition of Glycogen Synthase Kinase‐3β Activity"

    Article Title: Follistatin‐Like 3 Enhances the Function of Endothelial Cells Derived from Pluripotent Stem Cells by Facilitating β‐Catenin Nuclear Translocation Through Inhibition of Glycogen Synthase Kinase‐3β Activity

    Journal: Stem Cells (Dayton, Ohio)

    doi: 10.1002/stem.2820

    Induced pluripotent stem cell differentiation toward endothelial cells. Human iPS Cells were differentiated using StemPro serum free media supplemented with BMP4, Activin A, fibroblast growth factor (FGF), and vascular endothelial growth factor (VEGF) for 5 days. The differentiated cells were seeded on collagen IV, while CD144 positive cells were magnetically sorted on day 6 using MicroBeads Kit (Miltenyi BIotec) and culturing in EGM‐2 media (LONZA). (A) : Images show morphology of iPS Cells (left panel) and of their differentiated EC counterparts (right panel) Scale bar: 50 μm. (B) : Real time polymerase chain reaction (PCR) data revealed significantly increased early and (C) late EC marker mRNA expression (Data is means ±SEM [ n = 3], *, p
    Figure Legend Snippet: Induced pluripotent stem cell differentiation toward endothelial cells. Human iPS Cells were differentiated using StemPro serum free media supplemented with BMP4, Activin A, fibroblast growth factor (FGF), and vascular endothelial growth factor (VEGF) for 5 days. The differentiated cells were seeded on collagen IV, while CD144 positive cells were magnetically sorted on day 6 using MicroBeads Kit (Miltenyi BIotec) and culturing in EGM‐2 media (LONZA). (A) : Images show morphology of iPS Cells (left panel) and of their differentiated EC counterparts (right panel) Scale bar: 50 μm. (B) : Real time polymerase chain reaction (PCR) data revealed significantly increased early and (C) late EC marker mRNA expression (Data is means ±SEM [ n = 3], *, p

    Techniques Used: Cell Differentiation, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Marker, Expressing

    27) Product Images from "Per2 Mutation Recapitulates the Vascular Phenotype of Diabetes in the Retina and Bone Marrow"

    Article Title: Per2 Mutation Recapitulates the Vascular Phenotype of Diabetes in the Retina and Bone Marrow

    Journal: Diabetes

    doi: 10.2337/db12-0172

    Decrease in VEGF receptor expression and upregulation of TGF-β-induced profibrotic gene expression in Per2 mutant retinas. There was no significant change in VEGFR1 ( A ) or VEGFR2 ( B ) expression in retinas of 4-month-old Per2 mutant mice; however, there was a dramatic decrease in mRNA expression of both receptor expressions in 12-month-old animals. VEGF expression remained unchanged in both groups ( C ). Per2 mutant retinas showed a profound increase in mRNA expression of TGF-β1 ( D ) and its downstream effectors PAI-1 ( E ), ID-1 ( F ), CTGF ( G ), and FN ( H ). The most prevalent change in mRNA expression was observed in 4-month-old Per2 mutant mice for TGF-β1, PAI-1, and FN, whereas ID-1 expression was significantly increased in 12-month-old Per2 mutant mice. CTGF mRNA was increased in both age groups. n = 7 for 4-month-old mice; n = 3 for 12-month-old mice; white bars, wild type; black bars, Per2 mutant.
    Figure Legend Snippet: Decrease in VEGF receptor expression and upregulation of TGF-β-induced profibrotic gene expression in Per2 mutant retinas. There was no significant change in VEGFR1 ( A ) or VEGFR2 ( B ) expression in retinas of 4-month-old Per2 mutant mice; however, there was a dramatic decrease in mRNA expression of both receptor expressions in 12-month-old animals. VEGF expression remained unchanged in both groups ( C ). Per2 mutant retinas showed a profound increase in mRNA expression of TGF-β1 ( D ) and its downstream effectors PAI-1 ( E ), ID-1 ( F ), CTGF ( G ), and FN ( H ). The most prevalent change in mRNA expression was observed in 4-month-old Per2 mutant mice for TGF-β1, PAI-1, and FN, whereas ID-1 expression was significantly increased in 12-month-old Per2 mutant mice. CTGF mRNA was increased in both age groups. n = 7 for 4-month-old mice; n = 3 for 12-month-old mice; white bars, wild type; black bars, Per2 mutant.

    Techniques Used: Expressing, Mutagenesis, Mouse Assay

    28) Product Images from "Enhanced expression of mRNA for nuclear factor ?B1 (p50) in CD34+ cells of the bone marrow in rheumatoid arthritis"

    Article Title: Enhanced expression of mRNA for nuclear factor ?B1 (p50) in CD34+ cells of the bone marrow in rheumatoid arthritis

    Journal: Arthritis Research & Therapy

    doi: 10.1186/ar1915

    Suppression of the production of matrix metalloproteinase (MMP)-1 and vascular endothelial growth factor (VEGF) by silencing nuclear factor (NF)κB1 mRNA in bone marrow CD34+ cells from patients with rheumatoid arthritis. Purified bone marrow CD34+ cells from 12 patients with rheumatoid arthritis were transfected with small interfering RNA (siRNA) for NFκB1 or a scrambled sequence control siRNA, after which the cells were further incubated in culture medium with stem cell factor (10 ng/ml), granulocyte-macrophage colony stimulating factor (1 ng/ml) and tumor necrosis factor-α (10 ng/ml) for 4 weeks with no medium changes. After the incubation, the supernatants were harvested and assayed for MMP-1 and VEGF by ELISA. Statistical significance was evaluated by Wilcoxon's signed rank test.
    Figure Legend Snippet: Suppression of the production of matrix metalloproteinase (MMP)-1 and vascular endothelial growth factor (VEGF) by silencing nuclear factor (NF)κB1 mRNA in bone marrow CD34+ cells from patients with rheumatoid arthritis. Purified bone marrow CD34+ cells from 12 patients with rheumatoid arthritis were transfected with small interfering RNA (siRNA) for NFκB1 or a scrambled sequence control siRNA, after which the cells were further incubated in culture medium with stem cell factor (10 ng/ml), granulocyte-macrophage colony stimulating factor (1 ng/ml) and tumor necrosis factor-α (10 ng/ml) for 4 weeks with no medium changes. After the incubation, the supernatants were harvested and assayed for MMP-1 and VEGF by ELISA. Statistical significance was evaluated by Wilcoxon's signed rank test.

    Techniques Used: Purification, Transfection, Small Interfering RNA, Sequencing, Incubation, Enzyme-linked Immunosorbent Assay

    Suppression of the production of matrix metalloproteinase (MMP)-1 and vascular endothelial growth factor (VEGF) by silencing nuclear factor (NF)κB1 mRNA in bone marrow CD34+ cells from patients with rheumatoid arthritis. Purified bone marrow CD34+ cells from 12 patients with rheumatoid arthritis were transfected with small interfering RNA (siRNA) for NFκB1 or a scrambled sequence control siRNA, after which the cells were further incubated in culture medium with stem cell factor (10 ng/ml), granulocyte-macrophage colony stimulating factor (1 ng/ml) and tumor necrosis factor-α (10 ng/ml) for 4 weeks with no medium changes. After the incubation, the supernatants were harvested and assayed for MMP-1, VEGF and β 2 -microglobulin (β 2 MG) by ELISA. Statistical significance was evaluated by Wilcoxon's signed rank test.
    Figure Legend Snippet: Suppression of the production of matrix metalloproteinase (MMP)-1 and vascular endothelial growth factor (VEGF) by silencing nuclear factor (NF)κB1 mRNA in bone marrow CD34+ cells from patients with rheumatoid arthritis. Purified bone marrow CD34+ cells from 12 patients with rheumatoid arthritis were transfected with small interfering RNA (siRNA) for NFκB1 or a scrambled sequence control siRNA, after which the cells were further incubated in culture medium with stem cell factor (10 ng/ml), granulocyte-macrophage colony stimulating factor (1 ng/ml) and tumor necrosis factor-α (10 ng/ml) for 4 weeks with no medium changes. After the incubation, the supernatants were harvested and assayed for MMP-1, VEGF and β 2 -microglobulin (β 2 MG) by ELISA. Statistical significance was evaluated by Wilcoxon's signed rank test.

    Techniques Used: Purification, Transfection, Small Interfering RNA, Sequencing, Incubation, Enzyme-linked Immunosorbent Assay

    29) Product Images from "Cultured Human Epidermis Combined With Meshed Skin Autografts Accelerates Epithelialization and Granulation Tissue Formation in a Rat Model"

    Article Title: Cultured Human Epidermis Combined With Meshed Skin Autografts Accelerates Epithelialization and Granulation Tissue Formation in a Rat Model

    Journal: Annals of Plastic Surgery

    doi: 10.1097/SAP.0000000000001058

    Growth factor release by hCE. The release of bFGF, IL-1α, IL-1β, IL-6, PDGF-AA, TGF-α, TGF-β1, VEGF, and KGF by hCE into medium after 24-hour culture. The vertical axis represents logarithmic scale. N.D., not detected.
    Figure Legend Snippet: Growth factor release by hCE. The release of bFGF, IL-1α, IL-1β, IL-6, PDGF-AA, TGF-α, TGF-β1, VEGF, and KGF by hCE into medium after 24-hour culture. The vertical axis represents logarithmic scale. N.D., not detected.

    Techniques Used:

    30) Product Images from "(Pro)renin Receptor Triggers Distinct Angiotensin II-Independent Extracellular Matrix Remodeling and Deterioration of Cardiac Function"

    Article Title: (Pro)renin Receptor Triggers Distinct Angiotensin II-Independent Extracellular Matrix Remodeling and Deterioration of Cardiac Function

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0041404

    Effect of intramyocardial (P)RR gene delivery on cardiac gene expression without or with the losartan (Los) treatment. A, atrial natriuretic peptide (ANP) mRNA, B, β-myosin heavy chain (β-MHC) mRNA, C, skeletal α-actin mRNA, D, α-myosin heavy chain (α-MHC), E, cardiac α-actin mRNA, F, sarcoplasmic reticulum Ca 2+ ATPase (SERCA2) mRNA, G, vascular endothelial growth factor (VEGF) mRNA at 2 weeks and H, fibroblast growth factor-2 (FGF-2) mRNA levels at 1 week. The results are expressed as mean±SEM (n = 8 to 10). * P
    Figure Legend Snippet: Effect of intramyocardial (P)RR gene delivery on cardiac gene expression without or with the losartan (Los) treatment. A, atrial natriuretic peptide (ANP) mRNA, B, β-myosin heavy chain (β-MHC) mRNA, C, skeletal α-actin mRNA, D, α-myosin heavy chain (α-MHC), E, cardiac α-actin mRNA, F, sarcoplasmic reticulum Ca 2+ ATPase (SERCA2) mRNA, G, vascular endothelial growth factor (VEGF) mRNA at 2 weeks and H, fibroblast growth factor-2 (FGF-2) mRNA levels at 1 week. The results are expressed as mean±SEM (n = 8 to 10). * P

    Techniques Used: Expressing, Aqueous Normal-phase Chromatography

    31) Product Images from "EFFECT OF RHO-KINASE INHIBITOR, Y27632, ON PORCINE CORNEAL ENDOTHELIAL CELL CULTURE, INFLAMMATION AND IMMUNE REGULATION"

    Article Title: EFFECT OF RHO-KINASE INHIBITOR, Y27632, ON PORCINE CORNEAL ENDOTHELIAL CELL CULTURE, INFLAMMATION AND IMMUNE REGULATION

    Journal: Ocular immunology and inflammation

    doi: 10.3109/09273948.2015.1056534

    Suppression of inflammatory chemokines from pCECs (A) MCP-1 and VEGF mRNA levels in pCECs with/without 10μM Y27632 were measured by real-time PCR after hTNF-α activation. Although there were increases in MCP-1 and VEGF mRNA in pCECs after activation, Y27632 significantly reduced MCP-1 and VEGF mRNA expression after 6h of activation (43% and 37% reduction respectively, *p
    Figure Legend Snippet: Suppression of inflammatory chemokines from pCECs (A) MCP-1 and VEGF mRNA levels in pCECs with/without 10μM Y27632 were measured by real-time PCR after hTNF-α activation. Although there were increases in MCP-1 and VEGF mRNA in pCECs after activation, Y27632 significantly reduced MCP-1 and VEGF mRNA expression after 6h of activation (43% and 37% reduction respectively, *p

    Techniques Used: Real-time Polymerase Chain Reaction, Activation Assay, Expressing

    32) Product Images from "Level of Fatty Acid Binding Protein 5 (FABP5) Is Increased in Sputum of Allergic Asthmatics and Links to Airway Remodeling and Inflammation"

    Article Title: Level of Fatty Acid Binding Protein 5 (FABP5) Is Increased in Sputum of Allergic Asthmatics and Links to Airway Remodeling and Inflammation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0127003

    Immunological confirmation of findings. 2D-DIGE gel spot abundances for the FABP5 of the 21 induced sputum samples (A). Western blot analysis for 138 induced sputum samples (B) and for 158 nasal lavage fluid samples (C) with FABP5 antibody. ELISA results for vascular endothelial growth factor (VEGF) (D) and cysteinyl leukotriene (CysLT) (E) measurements from 90 nasal lavage fluid samples. The figures show mean with standard error of the mean. * p
    Figure Legend Snippet: Immunological confirmation of findings. 2D-DIGE gel spot abundances for the FABP5 of the 21 induced sputum samples (A). Western blot analysis for 138 induced sputum samples (B) and for 158 nasal lavage fluid samples (C) with FABP5 antibody. ELISA results for vascular endothelial growth factor (VEGF) (D) and cysteinyl leukotriene (CysLT) (E) measurements from 90 nasal lavage fluid samples. The figures show mean with standard error of the mean. * p

    Techniques Used: Western Blot, Enzyme-linked Immunosorbent Assay

    Related Articles

    SYBR Green Assay:

    Article Title: Exosome Delivered Anticancer Drugs Across the Blood-Brain Barrier for Brain Cancer Therapy in Danio Rerio
    Article Snippet: .. Exosome isolation solution, RNeasy Mini Kit, vascular endothelial growth factor (VEGF) forward and reverse primers, iTaq™ Universal SYBR® Green One-Step Kit, and biological agents were purchased from Life Technologies (Grand Island, NY, USA). .. CD9, CD63, and CD81 antibodies, ELISA kits, and exosome-depleted FBS were obtained from System Biosciences Inc. (Mountain View, CA, USA).

    Article Title: Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress
    Article Snippet: .. The mRNA expression levels of the growth factors, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), interleukin-1β (IL-1β ), stromal-derived factor-1 (SDF-1), the chemokine receptor CXCR4, and normalizing housekeeping genes GUSB and RLP27 (see for sequence information) were measured by real-time reverse transcription polymerase chain reaction (RT-PCR) (StepOnePlus, Applied Biosystems) using SYBR® Green qPCR Mastermix (Qiagen). .. The cycling RT-PCR conditions were as follows: 10 min at 95°C, 40 cycles for 10 s at 95°C, 15 s at 60°C, followed by gradient stage from 60 to 95°C to obtain a melting curve.

    Article Title: Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress
    Article Snippet: .. The mRNA expression levels of the growth factors, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), interleukin-1 β (IL-1 β ), stromal-derived factor-1 (SDF-1), the chemokine receptor CXCR4, and normalizing housekeeping genes GUSB and RLP27 (see for sequence information) were measured by real-time reverse transcription polymerase chain reaction (RT-PCR) (StepOnePlus, Applied Biosystems) using SYBR® Green qPCR Mastermix (Qiagen). .. The cycling RT-PCR conditions were as follows: 10 min at 95°C, 40 cycles for 10 s at 95°C, 15 s at 60°C, followed by gradient stage from 60 to 95°C to obtain a melting curve.

    Isolation:

    Article Title: Exosome Delivered Anticancer Drugs Across the Blood-Brain Barrier for Brain Cancer Therapy in Danio Rerio
    Article Snippet: .. Exosome isolation solution, RNeasy Mini Kit, vascular endothelial growth factor (VEGF) forward and reverse primers, iTaq™ Universal SYBR® Green One-Step Kit, and biological agents were purchased from Life Technologies (Grand Island, NY, USA). .. CD9, CD63, and CD81 antibodies, ELISA kits, and exosome-depleted FBS were obtained from System Biosciences Inc. (Mountain View, CA, USA).

    Enzyme-linked Immunosorbent Assay:

    Article Title: Xanthohumol Impairs Human Prostate Cancer Cell Growth and Invasion and Diminishes the Incidence and Progression of Advanced Tumors in TRAMP Mice
    Article Snippet: .. Vascular endothelial growth factor (VEGF) released into the media by PCa cells was measured using a commercial human ELISA kit (BioSource, Invitrogen; Life Technologies, Carlsbad, CA, USA). .. Use of TRAMP C57BL/6 mice for the studies described herein was approved by the Institutional Animal Care and Use Committee and complied with the Guide for the Care and Use of Laboratory Animals ( ).

    Article Title: Adipose tissue-derived extracellular fraction characterization: biological and clinical considerations in regenerative medicine
    Article Snippet: .. ELISA assay Growth factor and bioactive molecule concentrations in AT-Ex and plasma were measured using commercially available enzyme-linked immunosorbent assay (ELISA) kits according to the manufacturer’s instructions: basic fibroblast growth factor (bFGF; RayBio, Inc., Norcross, GA, USA), epidermal growth factor (EGF), erythropoietin (Epo), granulocyte/macrophage colony-stimulating factor (GM-CSF), and vascular endothelial growth factor (VEGF)-A (eBioscience, Inc. San Diego, CA, USA), keratinocyte growth factor (KGF; Cohesion Biosciences Ltd., London, UK), wingless type (Wnt)3a and Wnt10b (Cloud-Clone Corp., Katy, TX, USA), hepatocyte growth factor (HGF), stem cell factor (SCF), and α-melanocyte stimulating hormone (α-MSH; Cusabio Technology LLC, Baltimore, MD, USA). .. Matrix metalloproteinase (MMP)2 (4Abio Co. Ltd., Slough, UK) and Elastin (Elabiosciences, Houston, TX, USA) release in culture medium by photodamaged cells was measured after 2 weeks of treatment and results were normalized against protein concentration.

    Sequencing:

    Article Title: Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress
    Article Snippet: .. The mRNA expression levels of the growth factors, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), interleukin-1β (IL-1β ), stromal-derived factor-1 (SDF-1), the chemokine receptor CXCR4, and normalizing housekeeping genes GUSB and RLP27 (see for sequence information) were measured by real-time reverse transcription polymerase chain reaction (RT-PCR) (StepOnePlus, Applied Biosystems) using SYBR® Green qPCR Mastermix (Qiagen). .. The cycling RT-PCR conditions were as follows: 10 min at 95°C, 40 cycles for 10 s at 95°C, 15 s at 60°C, followed by gradient stage from 60 to 95°C to obtain a melting curve.

    Article Title: Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress
    Article Snippet: .. The mRNA expression levels of the growth factors, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), interleukin-1 β (IL-1 β ), stromal-derived factor-1 (SDF-1), the chemokine receptor CXCR4, and normalizing housekeeping genes GUSB and RLP27 (see for sequence information) were measured by real-time reverse transcription polymerase chain reaction (RT-PCR) (StepOnePlus, Applied Biosystems) using SYBR® Green qPCR Mastermix (Qiagen). .. The cycling RT-PCR conditions were as follows: 10 min at 95°C, 40 cycles for 10 s at 95°C, 15 s at 60°C, followed by gradient stage from 60 to 95°C to obtain a melting curve.

    Expressing:

    Article Title: Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress
    Article Snippet: .. The mRNA expression levels of the growth factors, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), interleukin-1β (IL-1β ), stromal-derived factor-1 (SDF-1), the chemokine receptor CXCR4, and normalizing housekeeping genes GUSB and RLP27 (see for sequence information) were measured by real-time reverse transcription polymerase chain reaction (RT-PCR) (StepOnePlus, Applied Biosystems) using SYBR® Green qPCR Mastermix (Qiagen). .. The cycling RT-PCR conditions were as follows: 10 min at 95°C, 40 cycles for 10 s at 95°C, 15 s at 60°C, followed by gradient stage from 60 to 95°C to obtain a melting curve.

    Article Title: Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress
    Article Snippet: .. The mRNA expression levels of the growth factors, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), interleukin-1 β (IL-1 β ), stromal-derived factor-1 (SDF-1), the chemokine receptor CXCR4, and normalizing housekeeping genes GUSB and RLP27 (see for sequence information) were measured by real-time reverse transcription polymerase chain reaction (RT-PCR) (StepOnePlus, Applied Biosystems) using SYBR® Green qPCR Mastermix (Qiagen). .. The cycling RT-PCR conditions were as follows: 10 min at 95°C, 40 cycles for 10 s at 95°C, 15 s at 60°C, followed by gradient stage from 60 to 95°C to obtain a melting curve.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress
    Article Snippet: .. The mRNA expression levels of the growth factors, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), interleukin-1β (IL-1β ), stromal-derived factor-1 (SDF-1), the chemokine receptor CXCR4, and normalizing housekeeping genes GUSB and RLP27 (see for sequence information) were measured by real-time reverse transcription polymerase chain reaction (RT-PCR) (StepOnePlus, Applied Biosystems) using SYBR® Green qPCR Mastermix (Qiagen). .. The cycling RT-PCR conditions were as follows: 10 min at 95°C, 40 cycles for 10 s at 95°C, 15 s at 60°C, followed by gradient stage from 60 to 95°C to obtain a melting curve.

    Article Title: Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress
    Article Snippet: .. The mRNA expression levels of the growth factors, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), interleukin-1 β (IL-1 β ), stromal-derived factor-1 (SDF-1), the chemokine receptor CXCR4, and normalizing housekeeping genes GUSB and RLP27 (see for sequence information) were measured by real-time reverse transcription polymerase chain reaction (RT-PCR) (StepOnePlus, Applied Biosystems) using SYBR® Green qPCR Mastermix (Qiagen). .. The cycling RT-PCR conditions were as follows: 10 min at 95°C, 40 cycles for 10 s at 95°C, 15 s at 60°C, followed by gradient stage from 60 to 95°C to obtain a melting curve.

    Real-time Polymerase Chain Reaction:

    Article Title: Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress
    Article Snippet: .. The mRNA expression levels of the growth factors, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), interleukin-1β (IL-1β ), stromal-derived factor-1 (SDF-1), the chemokine receptor CXCR4, and normalizing housekeeping genes GUSB and RLP27 (see for sequence information) were measured by real-time reverse transcription polymerase chain reaction (RT-PCR) (StepOnePlus, Applied Biosystems) using SYBR® Green qPCR Mastermix (Qiagen). .. The cycling RT-PCR conditions were as follows: 10 min at 95°C, 40 cycles for 10 s at 95°C, 15 s at 60°C, followed by gradient stage from 60 to 95°C to obtain a melting curve.

    Article Title: Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress
    Article Snippet: .. The mRNA expression levels of the growth factors, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), interleukin-1 β (IL-1 β ), stromal-derived factor-1 (SDF-1), the chemokine receptor CXCR4, and normalizing housekeeping genes GUSB and RLP27 (see for sequence information) were measured by real-time reverse transcription polymerase chain reaction (RT-PCR) (StepOnePlus, Applied Biosystems) using SYBR® Green qPCR Mastermix (Qiagen). .. The cycling RT-PCR conditions were as follows: 10 min at 95°C, 40 cycles for 10 s at 95°C, 15 s at 60°C, followed by gradient stage from 60 to 95°C to obtain a melting curve.

    Recombinant:

    Article Title: CYR61, a product of a growth factor-inducible immediate early gene, promotes angiogenesis and tumor growth
    Article Snippet: .. Recombinant human basic fibroblast growth factor (bFGF) was obtained from GIBCO/BRL; vascular endothelial growth factor (VEGF) was obtained from R & D Systems. .. Recombinant human CYR61 protein was purified from serum-free conditioned media of Sf-9 insect cells programmed for synthesis of the CYR61 protein via a baculovirus expression vector ( , ).

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    • vegfr 1  (Thermo Fisher)
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    Thermo Fisher vegfr 1
    Influence of <t>VEGFR‐1</t> silencing on the susceptibility to vemurafenib of M14‐VR and M14 cells. A and B, M14‐VR (A) or M14 (B) cells (1000/well) were seeded into 96‐well plates and the day after transfected with 10 nmol/L siVEGFR‐1 or siCTR and treated with graded concentrations of vemurafenib. After 5 d of culture, cell growth was analysed by MTS assay. Data are the mean of three independent experiments. Statistical analysis by two‐tailed Student's t test: * P
    Vegfr 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    vegf a  (Thermo Fisher)
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    Thermo Fisher vegf a
    KAT7 depletion disrupts endothelial cell activity. A, representative images of matrigel assays on control and KAT7-depleted ECs. The proportion of isolated and network branches, number of branch points, and total number of branches in matrigel assays on KAT7 siRNAs versus control siRNA-treated <t>HUVECs</t> are quantified. Data represent mean ± S.E. ( error bars ) of three replicates in a representative experiment of the matrigel assay. Biological replicates show similar results ( n = 3). * denotes a statistical significance of p ≤ 0.05 compared with control siRNA. B, representative images of wound healing assay on control and KAT7-depleted ECs with the quantification of the scratch area recovered. Data represent mean ± S.E. ( error bars ) of three independent wound healing experiments. * denotes a statistical significance of p ≤ 0.05 compared with control siRNA. Representative composite images of the matrigel and wound-healing assays were acquired from a Zeiss/AxioObserver.Z1 microscope (magnification: ×10, numerical aperture: 0.3) equipped with an EC Plan-Neofluar objective lens and environmental control (37 °C and 5% CO 2 with humidity) using a Hamamatsu ORCA-R2 camera. Composite images were extracted with the Zen Blue 2012 software. The black dashed lines in the composite images of the matrigel assay and gray dashed lines in the composite images of the wound-healing assays denote the splice sites between stitched individual images. C, representative images of cell migration assays conducted on control and KAT7-depleted ECs in the presence of <t>VEGF-A</t> 165 . The number of migrated cells is quantified. Representative images of the cell migration assays were acquired from an Olympus upright BX50 microscope (magnification: ×10, numerical aperture: 0.2) equipped with a DP72 camera. Images were extracted using the CellSens software. Data represent mean ± S.E. ( error bars ) of four independent experiments. * denotes a statistical significance of p ≤ 0.05 compared with control siRNA in the absence of VEGF-A 165 . D, representative immunoblot of VEGFR-2 tyrosine 1175 phosphorylation ( phospho-VEGFR-2 Y1175 ), VEGFR-2, KAT7, and α-tubulin in control and KAT7-depleted ECs treated with VEGF-A 165 for 0, 3, 5, 10, and 30 min. Gaps that represent empty single lanes between the control and KAT7-depleted samples in the P-VEGFR2 Tyr-1175, KAT7, and α-tubulin immunoblots were removed from the images. Quantification of VEGFR-2 tyrosine 1175 ( Y1175 ) phosphorylation immunoblots after normalizing for VEGFR-2 expression. Data represent mean ± S.E. ( error bars ) of three independent experiments. * denotes a statistical significance of p≤0.05 compared with control siRNA.
    Vegf A, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 106 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    vegfr 2  (Thermo Fisher)
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    VEGF-induced activation of c-Src, Akt, but not <t>VEGFR-2,</t> PLCγ-1 and ERK1/2 requires NADPH oxidase-derived ROS. (A) Protein extracts from HCAEC transfected with control siRNA (Scram-si) or si-p47 phox were subject to Western blots as described in Materials and methods . Serum-starved HCAEC were treated with VEGF (50 ng/ml) for the times indicated. Membranes were sequentially blotted, stripped and re-probed with the phospho-specific antibodies as shown. Blots shown are representative of three independent experiments. (B) Same as in (A) except, the membranes were probed for phosphorylation of Y1175 VEGFR-2, Y783 PLCγ-1 and p42/44 ERK1/2. Anti-β-actin antibody was used as loading control.
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    Influence of VEGFR‐1 silencing on the susceptibility to vemurafenib of M14‐VR and M14 cells. A and B, M14‐VR (A) or M14 (B) cells (1000/well) were seeded into 96‐well plates and the day after transfected with 10 nmol/L siVEGFR‐1 or siCTR and treated with graded concentrations of vemurafenib. After 5 d of culture, cell growth was analysed by MTS assay. Data are the mean of three independent experiments. Statistical analysis by two‐tailed Student's t test: * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Role of VEGFR‐1 in melanoma acquired resistance to the BRAF inhibitor vemurafenib, et al. Role of VEGFR‐1 in melanoma acquired resistance to the BRAF inhibitor vemurafenib

    doi: 10.1111/jcmm.14755

    Figure Lengend Snippet: Influence of VEGFR‐1 silencing on the susceptibility to vemurafenib of M14‐VR and M14 cells. A and B, M14‐VR (A) or M14 (B) cells (1000/well) were seeded into 96‐well plates and the day after transfected with 10 nmol/L siVEGFR‐1 or siCTR and treated with graded concentrations of vemurafenib. After 5 d of culture, cell growth was analysed by MTS assay. Data are the mean of three independent experiments. Statistical analysis by two‐tailed Student's t test: * P

    Article Snippet: Interestingly, VEGFR‐1 blockade by D16F7 mAb reduces ECM invasion triggered by VEGF‐A and PlGF, supporting the hypothesis that up‐regulation of VEGFR‐1 might contribute to tumour progression and spreading of melanoma after acquisition of a drug‐resistant phenotype.

    Techniques: Transfection, MTS Assay, Two Tailed Test

    Expression of VEGFR‐1 in melanoma cells with proliferative or invasive phenotypes and inhibitory effect of the anti‐VEGFR‐1 mAb D16F7 on ECM invasion by M14‐VR melanoma cells in response to PlGF or VEGF‐A. A, HOPP analysis based on VEGFR‐1 expression levels was carried out using gene expression data sets including 189 melanoma cell lines and short‐term cultures, of which 100 are characterized by a proliferative phenotype and 89 by an invasive phenotype. 33 Mean VEGFR‐1 transcript levels for proliferative (PRO) melanomas were compared with those of invasive melanomas (INV) and expressed as normalized signal intensity. Analysis of the 222033_s_at probeset for VEGFR‐1:3.9‐fold significant difference; statistical analysis by two‐tailed Student's t test: *** P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Role of VEGFR‐1 in melanoma acquired resistance to the BRAF inhibitor vemurafenib, et al. Role of VEGFR‐1 in melanoma acquired resistance to the BRAF inhibitor vemurafenib

    doi: 10.1111/jcmm.14755

    Figure Lengend Snippet: Expression of VEGFR‐1 in melanoma cells with proliferative or invasive phenotypes and inhibitory effect of the anti‐VEGFR‐1 mAb D16F7 on ECM invasion by M14‐VR melanoma cells in response to PlGF or VEGF‐A. A, HOPP analysis based on VEGFR‐1 expression levels was carried out using gene expression data sets including 189 melanoma cell lines and short‐term cultures, of which 100 are characterized by a proliferative phenotype and 89 by an invasive phenotype. 33 Mean VEGFR‐1 transcript levels for proliferative (PRO) melanomas were compared with those of invasive melanomas (INV) and expressed as normalized signal intensity. Analysis of the 222033_s_at probeset for VEGFR‐1:3.9‐fold significant difference; statistical analysis by two‐tailed Student's t test: *** P

    Article Snippet: Interestingly, VEGFR‐1 blockade by D16F7 mAb reduces ECM invasion triggered by VEGF‐A and PlGF, supporting the hypothesis that up‐regulation of VEGFR‐1 might contribute to tumour progression and spreading of melanoma after acquisition of a drug‐resistant phenotype.

    Techniques: Expressing, Two Tailed Test

    Enforced VEGFR‐1 expression in M14 melanoma cells increases invasiveness and reduces sensitivity to vemurafenib. A, VEGFR‐1 protein levels in M14 cells transfected with control (M14‐C) or VEGFR‐1 expressing (M14‐MF5) vectors were analysed by immunoblotting using antibodies against human VEGFR‐1 or β‐tubulin as loading control. The VEGFR‐1 protein has an expected molecular weight of 150 kD for the unmodified polypeptide and of 180‐185 kD for the glycosylated mature form. Positive control (Pos. CTR): glioblastoma cells transfected with the pBLAS49.2/VEGFR‐1 plasmid overexpressing the receptor. 42 B, Western blot of phosphorylated Erk (pErk) and total Erk in M14, M14‐C and M14‐MF5 cells; β‐tubulin (β‐Tub) was used as loading control. Histogram represents the densitometric quantification of band intensities, expressed as pErk/Erk ratio relative to M14 non transfected cells, after normalization for β‐tubulin expression. Normalized pErk1/Erk protein ratio in M14 cells was considered equal to 1.Data are the mean ± SD of three independent experiments. Statistical analysis by two‐tailed Student's t test: *** P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Role of VEGFR‐1 in melanoma acquired resistance to the BRAF inhibitor vemurafenib, et al. Role of VEGFR‐1 in melanoma acquired resistance to the BRAF inhibitor vemurafenib

    doi: 10.1111/jcmm.14755

    Figure Lengend Snippet: Enforced VEGFR‐1 expression in M14 melanoma cells increases invasiveness and reduces sensitivity to vemurafenib. A, VEGFR‐1 protein levels in M14 cells transfected with control (M14‐C) or VEGFR‐1 expressing (M14‐MF5) vectors were analysed by immunoblotting using antibodies against human VEGFR‐1 or β‐tubulin as loading control. The VEGFR‐1 protein has an expected molecular weight of 150 kD for the unmodified polypeptide and of 180‐185 kD for the glycosylated mature form. Positive control (Pos. CTR): glioblastoma cells transfected with the pBLAS49.2/VEGFR‐1 plasmid overexpressing the receptor. 42 B, Western blot of phosphorylated Erk (pErk) and total Erk in M14, M14‐C and M14‐MF5 cells; β‐tubulin (β‐Tub) was used as loading control. Histogram represents the densitometric quantification of band intensities, expressed as pErk/Erk ratio relative to M14 non transfected cells, after normalization for β‐tubulin expression. Normalized pErk1/Erk protein ratio in M14 cells was considered equal to 1.Data are the mean ± SD of three independent experiments. Statistical analysis by two‐tailed Student's t test: *** P

    Article Snippet: Interestingly, VEGFR‐1 blockade by D16F7 mAb reduces ECM invasion triggered by VEGF‐A and PlGF, supporting the hypothesis that up‐regulation of VEGFR‐1 might contribute to tumour progression and spreading of melanoma after acquisition of a drug‐resistant phenotype.

    Techniques: Expressing, Transfection, Molecular Weight, Positive Control, Plasmid Preparation, Western Blot, Two Tailed Test

    Influence of VEGFR‐1 silencing on the acquisition of resistance to vemurafenib by A375 cells. A, A375 cells were transfected with 10 nmol/L siVEGFR‐1 or siCTR and after three days total RNA was extracted and membrane VEGFR‐1 (mVEGFR‐1) transcript levels were assessed by qRT‐PCR analysis. Data are the mean of three independent determinations. Statistical analysis by two‐tailed Student's t test: *** P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Role of VEGFR‐1 in melanoma acquired resistance to the BRAF inhibitor vemurafenib, et al. Role of VEGFR‐1 in melanoma acquired resistance to the BRAF inhibitor vemurafenib

    doi: 10.1111/jcmm.14755

    Figure Lengend Snippet: Influence of VEGFR‐1 silencing on the acquisition of resistance to vemurafenib by A375 cells. A, A375 cells were transfected with 10 nmol/L siVEGFR‐1 or siCTR and after three days total RNA was extracted and membrane VEGFR‐1 (mVEGFR‐1) transcript levels were assessed by qRT‐PCR analysis. Data are the mean of three independent determinations. Statistical analysis by two‐tailed Student's t test: *** P

    Article Snippet: Interestingly, VEGFR‐1 blockade by D16F7 mAb reduces ECM invasion triggered by VEGF‐A and PlGF, supporting the hypothesis that up‐regulation of VEGFR‐1 might contribute to tumour progression and spreading of melanoma after acquisition of a drug‐resistant phenotype.

    Techniques: Transfection, Quantitative RT-PCR, Two Tailed Test

    Characterization of human melanoma cell lines sensitive or resistant to vemurafenib for the production of VEGF‐A and PlGF and expression of VEGFR‐1. VEGF‐A (A) and PlGF (B) secretion was quantified by ELISA (mean ± SD; n = 3). Each value represents the arithmetic mean of three independent experiments performed with triplicate samples. C, The expression of VEGFR‐1 transcript was assessed by qRT‐PCR analysis utilizing the human melanoma GR‐Mel cell line as positive control. The results are expressed as relative mRNA and are the mean ± SD of three (A375 lines) or two (M14 lines) independent determinations with duplicate samples. Data were referred to the VEGFR‐1 negative M14 bulk cell line, to which the arbitrary value of 1 was assigned. Statistical analysis by two‐tailed Student's t test: resistant vs sensitive cells: *** P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Role of VEGFR‐1 in melanoma acquired resistance to the BRAF inhibitor vemurafenib, et al. Role of VEGFR‐1 in melanoma acquired resistance to the BRAF inhibitor vemurafenib

    doi: 10.1111/jcmm.14755

    Figure Lengend Snippet: Characterization of human melanoma cell lines sensitive or resistant to vemurafenib for the production of VEGF‐A and PlGF and expression of VEGFR‐1. VEGF‐A (A) and PlGF (B) secretion was quantified by ELISA (mean ± SD; n = 3). Each value represents the arithmetic mean of three independent experiments performed with triplicate samples. C, The expression of VEGFR‐1 transcript was assessed by qRT‐PCR analysis utilizing the human melanoma GR‐Mel cell line as positive control. The results are expressed as relative mRNA and are the mean ± SD of three (A375 lines) or two (M14 lines) independent determinations with duplicate samples. Data were referred to the VEGFR‐1 negative M14 bulk cell line, to which the arbitrary value of 1 was assigned. Statistical analysis by two‐tailed Student's t test: resistant vs sensitive cells: *** P

    Article Snippet: Interestingly, VEGFR‐1 blockade by D16F7 mAb reduces ECM invasion triggered by VEGF‐A and PlGF, supporting the hypothesis that up‐regulation of VEGFR‐1 might contribute to tumour progression and spreading of melanoma after acquisition of a drug‐resistant phenotype.

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Positive Control, Two Tailed Test

    Inhibitory effect of the anti‐VEGFR‐1 mAb D16F7 on ECM invasion by A375‐VR melanoma cells in response to PlGF. A, ECM invasion of A375‐VR cells (2x10 5 cells/chamber, 4 h incubation) induced by PlGF (50 ng/mL) in the absence or presence of 5 µg/mL D16F7 mAb. BSA, non‐stimulated cells. Histograms represent the arithmetic mean ± SD of invading cells/microscopic field from three independent experiments. Statistical analysis was performed by Kruskal‐Wallis followed by Dunn's test for multiple comparison: P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Role of VEGFR‐1 in melanoma acquired resistance to the BRAF inhibitor vemurafenib, et al. Role of VEGFR‐1 in melanoma acquired resistance to the BRAF inhibitor vemurafenib

    doi: 10.1111/jcmm.14755

    Figure Lengend Snippet: Inhibitory effect of the anti‐VEGFR‐1 mAb D16F7 on ECM invasion by A375‐VR melanoma cells in response to PlGF. A, ECM invasion of A375‐VR cells (2x10 5 cells/chamber, 4 h incubation) induced by PlGF (50 ng/mL) in the absence or presence of 5 µg/mL D16F7 mAb. BSA, non‐stimulated cells. Histograms represent the arithmetic mean ± SD of invading cells/microscopic field from three independent experiments. Statistical analysis was performed by Kruskal‐Wallis followed by Dunn's test for multiple comparison: P

    Article Snippet: Interestingly, VEGFR‐1 blockade by D16F7 mAb reduces ECM invasion triggered by VEGF‐A and PlGF, supporting the hypothesis that up‐regulation of VEGFR‐1 might contribute to tumour progression and spreading of melanoma after acquisition of a drug‐resistant phenotype.

    Techniques: Incubation

    KAT7 depletion disrupts endothelial cell activity. A, representative images of matrigel assays on control and KAT7-depleted ECs. The proportion of isolated and network branches, number of branch points, and total number of branches in matrigel assays on KAT7 siRNAs versus control siRNA-treated HUVECs are quantified. Data represent mean ± S.E. ( error bars ) of three replicates in a representative experiment of the matrigel assay. Biological replicates show similar results ( n = 3). * denotes a statistical significance of p ≤ 0.05 compared with control siRNA. B, representative images of wound healing assay on control and KAT7-depleted ECs with the quantification of the scratch area recovered. Data represent mean ± S.E. ( error bars ) of three independent wound healing experiments. * denotes a statistical significance of p ≤ 0.05 compared with control siRNA. Representative composite images of the matrigel and wound-healing assays were acquired from a Zeiss/AxioObserver.Z1 microscope (magnification: ×10, numerical aperture: 0.3) equipped with an EC Plan-Neofluar objective lens and environmental control (37 °C and 5% CO 2 with humidity) using a Hamamatsu ORCA-R2 camera. Composite images were extracted with the Zen Blue 2012 software. The black dashed lines in the composite images of the matrigel assay and gray dashed lines in the composite images of the wound-healing assays denote the splice sites between stitched individual images. C, representative images of cell migration assays conducted on control and KAT7-depleted ECs in the presence of VEGF-A 165 . The number of migrated cells is quantified. Representative images of the cell migration assays were acquired from an Olympus upright BX50 microscope (magnification: ×10, numerical aperture: 0.2) equipped with a DP72 camera. Images were extracted using the CellSens software. Data represent mean ± S.E. ( error bars ) of four independent experiments. * denotes a statistical significance of p ≤ 0.05 compared with control siRNA in the absence of VEGF-A 165 . D, representative immunoblot of VEGFR-2 tyrosine 1175 phosphorylation ( phospho-VEGFR-2 Y1175 ), VEGFR-2, KAT7, and α-tubulin in control and KAT7-depleted ECs treated with VEGF-A 165 for 0, 3, 5, 10, and 30 min. Gaps that represent empty single lanes between the control and KAT7-depleted samples in the P-VEGFR2 Tyr-1175, KAT7, and α-tubulin immunoblots were removed from the images. Quantification of VEGFR-2 tyrosine 1175 ( Y1175 ) phosphorylation immunoblots after normalizing for VEGFR-2 expression. Data represent mean ± S.E. ( error bars ) of three independent experiments. * denotes a statistical significance of p≤0.05 compared with control siRNA.

    Journal: The Journal of Biological Chemistry

    Article Title: Histone acetyltransferase 7 (KAT7)-dependent intragenic histone acetylation regulates endothelial cell gene regulation

    doi: 10.1074/jbc.RA117.001383

    Figure Lengend Snippet: KAT7 depletion disrupts endothelial cell activity. A, representative images of matrigel assays on control and KAT7-depleted ECs. The proportion of isolated and network branches, number of branch points, and total number of branches in matrigel assays on KAT7 siRNAs versus control siRNA-treated HUVECs are quantified. Data represent mean ± S.E. ( error bars ) of three replicates in a representative experiment of the matrigel assay. Biological replicates show similar results ( n = 3). * denotes a statistical significance of p ≤ 0.05 compared with control siRNA. B, representative images of wound healing assay on control and KAT7-depleted ECs with the quantification of the scratch area recovered. Data represent mean ± S.E. ( error bars ) of three independent wound healing experiments. * denotes a statistical significance of p ≤ 0.05 compared with control siRNA. Representative composite images of the matrigel and wound-healing assays were acquired from a Zeiss/AxioObserver.Z1 microscope (magnification: ×10, numerical aperture: 0.3) equipped with an EC Plan-Neofluar objective lens and environmental control (37 °C and 5% CO 2 with humidity) using a Hamamatsu ORCA-R2 camera. Composite images were extracted with the Zen Blue 2012 software. The black dashed lines in the composite images of the matrigel assay and gray dashed lines in the composite images of the wound-healing assays denote the splice sites between stitched individual images. C, representative images of cell migration assays conducted on control and KAT7-depleted ECs in the presence of VEGF-A 165 . The number of migrated cells is quantified. Representative images of the cell migration assays were acquired from an Olympus upright BX50 microscope (magnification: ×10, numerical aperture: 0.2) equipped with a DP72 camera. Images were extracted using the CellSens software. Data represent mean ± S.E. ( error bars ) of four independent experiments. * denotes a statistical significance of p ≤ 0.05 compared with control siRNA in the absence of VEGF-A 165 . D, representative immunoblot of VEGFR-2 tyrosine 1175 phosphorylation ( phospho-VEGFR-2 Y1175 ), VEGFR-2, KAT7, and α-tubulin in control and KAT7-depleted ECs treated with VEGF-A 165 for 0, 3, 5, 10, and 30 min. Gaps that represent empty single lanes between the control and KAT7-depleted samples in the P-VEGFR2 Tyr-1175, KAT7, and α-tubulin immunoblots were removed from the images. Quantification of VEGFR-2 tyrosine 1175 ( Y1175 ) phosphorylation immunoblots after normalizing for VEGFR-2 expression. Data represent mean ± S.E. ( error bars ) of three independent experiments. * denotes a statistical significance of p≤0.05 compared with control siRNA.

    Article Snippet: HUVECs were induced with VEGF-A for 3, 5, 10, and 30 min at a final concentration of 50 ng/ml after serum starvation for 1 h with Media 199 (Thermo Fisher Scientific) in the VEGF-A signal transduction studies.

    Techniques: Activity Assay, Isolation, Matrigel Assay, Wound Healing Assay, Microscopy, Software, Migration, Western Blot, Expressing

    Survivin silencing decreases the malignant phenotype of Ras G12V -ΙκΒαΜ-derived tumors. ( A ) Hematoxilin and Eosin staining of survivin siRNA-treated Ras G12V -ΙκΒαΜ-derived tumors; ( B ) Mitotic Index representing the number of cells undergoing mitosis over total cells. Scale bars = 200 µm; ( C ) HIF1α staining in survivin siRNA transfected NoLacZ or Ras G12V -ΙκΒαΜ tumors evaluated by immunofluorescence; ( D ) VEGF staining in survivin siRNA transfected NoLacZ or Ras G12V -ΙκΒαΜ tumors evaluated by immunofluorescence; ( E ) CD51 staining in survivin siRNA transfected NoLacZ or Ras G12V -ΙκΒαΜ tumors evaluated by immunofluoresce; ( F ) HIF1α ( upper panel) and CD51 ( lower panel) positive cell count assessed by ImageJ software (* 0.01

    Journal: International Journal of Molecular Sciences

    Article Title: Survivin Modulates Squamous Cell Carcinoma-Derived Stem-Like Cell Proliferation, Viability and Tumor Formation in Vivo

    doi: 10.3390/ijms17010089

    Figure Lengend Snippet: Survivin silencing decreases the malignant phenotype of Ras G12V -ΙκΒαΜ-derived tumors. ( A ) Hematoxilin and Eosin staining of survivin siRNA-treated Ras G12V -ΙκΒαΜ-derived tumors; ( B ) Mitotic Index representing the number of cells undergoing mitosis over total cells. Scale bars = 200 µm; ( C ) HIF1α staining in survivin siRNA transfected NoLacZ or Ras G12V -ΙκΒαΜ tumors evaluated by immunofluorescence; ( D ) VEGF staining in survivin siRNA transfected NoLacZ or Ras G12V -ΙκΒαΜ tumors evaluated by immunofluorescence; ( E ) CD51 staining in survivin siRNA transfected NoLacZ or Ras G12V -ΙκΒαΜ tumors evaluated by immunofluoresce; ( F ) HIF1α ( upper panel) and CD51 ( lower panel) positive cell count assessed by ImageJ software (* 0.01

    Article Snippet: Then, slides were incubated for 15 min with 0.5% bovine serum albumin and 5% goat serum, and for 60 min at 37 °C with the rabbit polyclonal anti-human Survivin antibody (1:50, Novus, Bloomington, MN, USA) or HIF1α (1:500, Novus, Bloomington, MN, USA), VEGF (1:50, Thermo Fisher Scientific, Waltham, MA, USA), CD51 (1:50, Abcam, Cambridge, UK).

    Techniques: Derivative Assay, Staining, Transfection, Immunofluorescence, Cell Counting, Software

    VEGF-induced activation of c-Src, Akt, but not VEGFR-2, PLCγ-1 and ERK1/2 requires NADPH oxidase-derived ROS. (A) Protein extracts from HCAEC transfected with control siRNA (Scram-si) or si-p47 phox were subject to Western blots as described in Materials and methods . Serum-starved HCAEC were treated with VEGF (50 ng/ml) for the times indicated. Membranes were sequentially blotted, stripped and re-probed with the phospho-specific antibodies as shown. Blots shown are representative of three independent experiments. (B) Same as in (A) except, the membranes were probed for phosphorylation of Y1175 VEGFR-2, Y783 PLCγ-1 and p42/44 ERK1/2. Anti-β-actin antibody was used as loading control.

    Journal: PLoS ONE

    Article Title: Direct Sensing of Endothelial Oxidants by Vascular Endothelial Growth Factor Receptor-2 and c-Src

    doi: 10.1371/journal.pone.0028454

    Figure Lengend Snippet: VEGF-induced activation of c-Src, Akt, but not VEGFR-2, PLCγ-1 and ERK1/2 requires NADPH oxidase-derived ROS. (A) Protein extracts from HCAEC transfected with control siRNA (Scram-si) or si-p47 phox were subject to Western blots as described in Materials and methods . Serum-starved HCAEC were treated with VEGF (50 ng/ml) for the times indicated. Membranes were sequentially blotted, stripped and re-probed with the phospho-specific antibodies as shown. Blots shown are representative of three independent experiments. (B) Same as in (A) except, the membranes were probed for phosphorylation of Y1175 VEGFR-2, Y783 PLCγ-1 and p42/44 ERK1/2. Anti-β-actin antibody was used as loading control.

    Article Snippet: Colocalization of VEGFR-2 with c-Src or EEA1-positive endosome by immunofluorescence HCAEC were plated on fibronectin coated glass-bottom chamber slides (Lab-Tek II, Thermo Scientific) and starved overnight in 0.2% FBS containing EBM-2.

    Techniques: Activation Assay, Derivative Assay, Transfection, Western Blot

    VEGF-induced interaction between VEGFR-2 and c-Src requires NADPH oxidase-derived ROS. (A) Co-immunoprecipitation (co-IP) assay using 1.2 mg protein lysates of HCAEC that were transfected with Scram-si or si-p47 phox and treated without or with VEGF (50 ng/ml for 5 min). IP was carried out using anti-VEGFR-2 antibody followed by immunoblotting using anti-c-Src (upper panel) and anti-VEGFR-2 (lower panel) antibodies. (B) Quantitative analyses of VEGFR-2-bound c-Src. Bar graphs show quantitative densitometric analysis of three independent experiments using NIH image J (-fold change expressed in mean ± S.E.M.). * p

    Journal: PLoS ONE

    Article Title: Direct Sensing of Endothelial Oxidants by Vascular Endothelial Growth Factor Receptor-2 and c-Src

    doi: 10.1371/journal.pone.0028454

    Figure Lengend Snippet: VEGF-induced interaction between VEGFR-2 and c-Src requires NADPH oxidase-derived ROS. (A) Co-immunoprecipitation (co-IP) assay using 1.2 mg protein lysates of HCAEC that were transfected with Scram-si or si-p47 phox and treated without or with VEGF (50 ng/ml for 5 min). IP was carried out using anti-VEGFR-2 antibody followed by immunoblotting using anti-c-Src (upper panel) and anti-VEGFR-2 (lower panel) antibodies. (B) Quantitative analyses of VEGFR-2-bound c-Src. Bar graphs show quantitative densitometric analysis of three independent experiments using NIH image J (-fold change expressed in mean ± S.E.M.). * p

    Article Snippet: Colocalization of VEGFR-2 with c-Src or EEA1-positive endosome by immunofluorescence HCAEC were plated on fibronectin coated glass-bottom chamber slides (Lab-Tek II, Thermo Scientific) and starved overnight in 0.2% FBS containing EBM-2.

    Techniques: Derivative Assay, Co-Immunoprecipitation Assay, Transfection

    c-Src and VEGFR-2 are oxidized in VEGF-treated HCAEC in the presence of NADPH oxidase-derived ROS. (A) Schematic presentation depicting cysteinyl-labeling assay to determine oxidative modification in intracellular proteins. Non-oxidized protein thiols are alkyalated by IAA, oxidized thiols are reduced back to SH-moiety by DTT and subsequently biotinylated by IAP. Biotinylated proteins are then pulled-down by streptavidin-agarose followed by Western blots. (B) Upper panel: cysteinyl labeling assay to Identify thiol oxidation of proteins in VEGF-treated (50 ng/ml for 2 mins) HCAEC lysates using biotinylated IAP probe. HCAEC were transfected with Scram-si or si-p47 phox as indicated. After cell lysis in the presence of IAA followed by DTT treatment and IAP labeling, 1.5 mg biotinylated protein lysates were subject to immunoprecipitation using Streptavidin-agarose beads and immunoblotted using anti-VEGFR-2 and anti-c-Src antibodies. Lower panel: Western blot for VEGFR-2 using 50 µg of parallel HCAEC lysates as loading control. (B) Quantitative analyses of oxidized VEGFR-2 (upper panel) and c-Src (lower panel). Bar graph shows quantitative densitometric analysis of three independent cysteinyl labeling assays (as in A) using NIH J image (-fold change expressed in mean ± S.E.M.). * p

    Journal: PLoS ONE

    Article Title: Direct Sensing of Endothelial Oxidants by Vascular Endothelial Growth Factor Receptor-2 and c-Src

    doi: 10.1371/journal.pone.0028454

    Figure Lengend Snippet: c-Src and VEGFR-2 are oxidized in VEGF-treated HCAEC in the presence of NADPH oxidase-derived ROS. (A) Schematic presentation depicting cysteinyl-labeling assay to determine oxidative modification in intracellular proteins. Non-oxidized protein thiols are alkyalated by IAA, oxidized thiols are reduced back to SH-moiety by DTT and subsequently biotinylated by IAP. Biotinylated proteins are then pulled-down by streptavidin-agarose followed by Western blots. (B) Upper panel: cysteinyl labeling assay to Identify thiol oxidation of proteins in VEGF-treated (50 ng/ml for 2 mins) HCAEC lysates using biotinylated IAP probe. HCAEC were transfected with Scram-si or si-p47 phox as indicated. After cell lysis in the presence of IAA followed by DTT treatment and IAP labeling, 1.5 mg biotinylated protein lysates were subject to immunoprecipitation using Streptavidin-agarose beads and immunoblotted using anti-VEGFR-2 and anti-c-Src antibodies. Lower panel: Western blot for VEGFR-2 using 50 µg of parallel HCAEC lysates as loading control. (B) Quantitative analyses of oxidized VEGFR-2 (upper panel) and c-Src (lower panel). Bar graph shows quantitative densitometric analysis of three independent cysteinyl labeling assays (as in A) using NIH J image (-fold change expressed in mean ± S.E.M.). * p

    Article Snippet: Colocalization of VEGFR-2 with c-Src or EEA1-positive endosome by immunofluorescence HCAEC were plated on fibronectin coated glass-bottom chamber slides (Lab-Tek II, Thermo Scientific) and starved overnight in 0.2% FBS containing EBM-2.

    Techniques: Derivative Assay, Labeling, Modification, Western Blot, Transfection, Lysis, Immunoprecipitation

    Proposed model: thiol oxidation may help propagate signal transduction from VEGFR-2 to downstream c-Src. (A) VEGF activation of VEGFR-2 and downstream PLCγ-1-ERK1/2 signaling pathway appears to be independent of ROS levels in ECs. (B) VEGF induces NADPH oxidase-derived ROS, which in turn oxidizes VEGFR-2 and c-Src. Thiol oxidation of these two tyrosine kinases appears to correlate with VEGF-induced activation of c-Src, and also with the sub-cellular colocalization and interaction between VEGFR-2 and c-Src. Dependence of VEGF-induced thiol oxidation and activation of c-Src on NADPH oxidase-derived ROS render downstream activation of PI3K-Akt redox-sensitive in HCAEC. In this model, VEGFR-2 and/or c-Src act as endothelial redox-sensors that determine whether downstream PI3K-Akt signaling pathway should be activated or not.

    Journal: PLoS ONE

    Article Title: Direct Sensing of Endothelial Oxidants by Vascular Endothelial Growth Factor Receptor-2 and c-Src

    doi: 10.1371/journal.pone.0028454

    Figure Lengend Snippet: Proposed model: thiol oxidation may help propagate signal transduction from VEGFR-2 to downstream c-Src. (A) VEGF activation of VEGFR-2 and downstream PLCγ-1-ERK1/2 signaling pathway appears to be independent of ROS levels in ECs. (B) VEGF induces NADPH oxidase-derived ROS, which in turn oxidizes VEGFR-2 and c-Src. Thiol oxidation of these two tyrosine kinases appears to correlate with VEGF-induced activation of c-Src, and also with the sub-cellular colocalization and interaction between VEGFR-2 and c-Src. Dependence of VEGF-induced thiol oxidation and activation of c-Src on NADPH oxidase-derived ROS render downstream activation of PI3K-Akt redox-sensitive in HCAEC. In this model, VEGFR-2 and/or c-Src act as endothelial redox-sensors that determine whether downstream PI3K-Akt signaling pathway should be activated or not.

    Article Snippet: Colocalization of VEGFR-2 with c-Src or EEA1-positive endosome by immunofluorescence HCAEC were plated on fibronectin coated glass-bottom chamber slides (Lab-Tek II, Thermo Scientific) and starved overnight in 0.2% FBS containing EBM-2.

    Techniques: Transduction, Activation Assay, Derivative Assay, Activated Clotting Time Assay

    VEGF induces subcellular co-localization of c-Src and internalized VEGFR-2 in an ROS-dependent manner. HCAEC transfected with control (Scram-si) (A) or si-p47 phox (B) were immunofluorescently double labeled for internalized VEGFR-2 (green) and c-Src (red). VEGFR-2 on HCAEC was labeled with single chain E-tagged antibody (scFvA7, Fitzerald) as described in Materials and methods . After incubation with VEGF (50 ng/ml for 10 min), in order to remove the antibody from the cell surface, cells were placed on ice and acid washed. In permeabilized and fixed HCAEC, VEGFR-2 was detected with an AlexaFluor488-conjugated secondary antibody and is shown in green. c-Src was labeled with AlexaFluor647-conjugated secondary antibody (red) and nuclei with DAPI (blue). (B) Bar graphs show image analysis for colocalization events using the NIH Image J plugin (as described in Materials and methods ). The graphs present the number of colocalization events normalized for the number of total VEGFR-2–positive immunofluorescence signals. Values are the mean of three experiments ± S.E.M., each containing numbers obtained from five random fields. * p

    Journal: PLoS ONE

    Article Title: Direct Sensing of Endothelial Oxidants by Vascular Endothelial Growth Factor Receptor-2 and c-Src

    doi: 10.1371/journal.pone.0028454

    Figure Lengend Snippet: VEGF induces subcellular co-localization of c-Src and internalized VEGFR-2 in an ROS-dependent manner. HCAEC transfected with control (Scram-si) (A) or si-p47 phox (B) were immunofluorescently double labeled for internalized VEGFR-2 (green) and c-Src (red). VEGFR-2 on HCAEC was labeled with single chain E-tagged antibody (scFvA7, Fitzerald) as described in Materials and methods . After incubation with VEGF (50 ng/ml for 10 min), in order to remove the antibody from the cell surface, cells were placed on ice and acid washed. In permeabilized and fixed HCAEC, VEGFR-2 was detected with an AlexaFluor488-conjugated secondary antibody and is shown in green. c-Src was labeled with AlexaFluor647-conjugated secondary antibody (red) and nuclei with DAPI (blue). (B) Bar graphs show image analysis for colocalization events using the NIH Image J plugin (as described in Materials and methods ). The graphs present the number of colocalization events normalized for the number of total VEGFR-2–positive immunofluorescence signals. Values are the mean of three experiments ± S.E.M., each containing numbers obtained from five random fields. * p

    Article Snippet: Colocalization of VEGFR-2 with c-Src or EEA1-positive endosome by immunofluorescence HCAEC were plated on fibronectin coated glass-bottom chamber slides (Lab-Tek II, Thermo Scientific) and starved overnight in 0.2% FBS containing EBM-2.

    Techniques: Transfection, Labeling, Incubation, Immunofluorescence