anti-matrix metalloproteinase mmp Search Results


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  • 94
    Millipore anti mmp 9
    Effects of postischemia-reperfusion (I/R) treatment with TMZ on the expressions of MMPs and TIMPs. Western blot analysis (a) and densitometric analysis (b–e) of the Western blot for MMP-2, -9 and TIMP-1, -2 at 5 d, and 8 w after I/R injury, respectively. Post I/R treatment with TMZ reduced the degree of I/R-induced increases in MMP-2 and <t>MMP-9</t> levels and further increased the levels of TIMP-1 and TIMP-2 at 5 d after renal I/R injury compared to the IRC group. At 8 w after I/R injury, MMP levels in the TMZ-treated groups were similar to those of the IRC groups, which were all significantly greater than those of the sham groups. TIMP levels in the TMZ-treated groups became also comparable to those of the IRC groups, which were significantly greater than those of the sham groups. TMZ = trimetazidine. Sham = rats not underwent I/R; IRC = rats underwent ischemia (45 min)-reperfusion (5 d or 8 w); TMZ S = rats treated with TMZ (3 mg/kg) upon reperfusion; TMZ D = rats treated with TMZ (3 mg/kg) once daily for 5 d or 8 w starting upon reperfusion. ∗ P
    Anti Mmp 9, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 321 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti mmp 9
    Mechanism of S. Typhimurium-induced intestinal damage during colitis. (A) Secretion of <t>MMP-9</t> and MMP-2 in the colons of mock (Con)- or S. Typhimurium (SaI)-infected Lcn2 −/− mice was analyzed by gelatin zymography using gelatin-agarose-purified PBS extracts (6–8 mice per group). (B) Quantitation of gelatinolytic activities from the zymogram in panel A and from another representative zymogram from independent experiments. (C) MMP-9 protein levels in the colons of mice from the respective groups were assessed by immunoblotting purified PBS extracts under non-reducing conditions. Flow-through from the purification was used as a loading control. (D) Quantitation of changes in protein levels from the immunoblot in panel C and from another representative blot from independent experiments. (E) MMP-9 expression levels in the colons of mock- or S. Typhimurium-infected Lcn2 +/+ and Lcn2 −/− mice were measured by real-time PCR. Error bars = ± standard error of the mean. *p
    Anti Mmp 9, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 828 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Santa Cruz Biotechnology anti mmp 9
    Effect of RPE treatment on levels of TGF-β1 and <t>MMP-9</t> in BAL. TGF-βand MMP-9 in BALwere measured by ELISA. Data are means ± SD from triplicate experiments. ** P
    Anti Mmp 9, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 773 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti mmp 9
    Effects of the ERK inhibitor (U0126) and baicalein on cell invasion and MMP-2, <t>MMP-9,</t> u-PA, TIMP-1 and TIMP-2 expression in MHCC97H cells. (A) Cells were pretreated with U0126 (10 μM) for 30 min and then incubated in the presence or absence of baicalein (10 μM) for 24 h. Cellular invasiveness was measured using the Boyden chamber invasion assay. (B) The percent invasion rate was expressed as a percentage of control. (C, D) MHCC97H cells were treated and then subjected to western blotting to analyze the protein levels of MMP-2, MMP-9, u-PA, TIMP-1 and TIMP-2. Values represent the means ± SD of three independent experiments performed in triplicate. * p
    Anti Mmp 9, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 599 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Santa Cruz Biotechnology anti mmp 2
    Curcumin retards activation of β-catenin/Slug pathway in breast cancer stem cells, thereby restoring E-cadherin. (A) β-catenin-associated E-cadherin was assayed by co-immunoprecipitation from cell lysates of MCF-7-derived 2° mammospheres with or without curcumin treatment using specific antibodies (left panel) or with normal human immunoglobulin G (IgG) as a negative control (right panel). To ensure comparable protein loading, 20% of supernatant from immunoprecipitation (IP) sample was subjected to determination of α-actin by Western blotting. (B) Western blotting was conducted to study the levels of total β-catenin and nuclear β-catenin in 2° mammospheres in presence or absence of curcumin exposure. (C) The relative nuclear expression of β-catenin in 2° mammospheres with or without curcumin treatment was visualized by immunofluorescence. (D, E) Under similar conditions, Western blotting was performed to study the expression levels of β-catenin target genes Cyclin-D1, c-Myc, Slug, Snail, Vimentin, <t>MMP-2</t> and MMP-9 in curcumin treated/untreated 2° mammospheres. (F) Protein expression of E-cadherin in 2° mammospheres with or without curcumin or Slug-cDNA (or both) were determined by Western blotting (left panel). The efficiency of transfection was determined by Western blot analysis (right panel). (G) Graphical quantifications of cell adhesion, spreading, three-dimensional (3D) invasion, and migration assays of MCF-7-derived 2° mammospheres with or without curcumin and Slug-cDNA treatment/transfection. (H) Transwell migration assay was performed under similar experimental conditions in T47D-derived 2° mammospheres. α-Actin/histone H1/glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal loading control. Data are presented as mean ± standard error of mean or representative of three independent experiments. Cont, control; Cur, curcumin.
    Anti Mmp 2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 488 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti mmp 2
    TLR2 blockade inhibits vascular inflammation in atherosclerotic arteries. <t>MMP-2</t> expression (A and B), NF-κB p65 phosphorylation (C), Stat3 phosphorylation (D) and the expression of the cytokines IL-6, TNF-α, and IL-10 (E) were reduced
    Anti Mmp 2, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 877 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mmp 9  (Abcam)
    98
    Abcam mmp 9
    p-AKT, MMP-2 and <t>MMP-9</t> are involved in PRL-3-mediated invasion and migration of gastric cancer cells. (A) Levels of PRL-3, AKT and p-AKT were analyzed in SGC7901 cells (control), SGC7901 cells transfected with empty vector, and plasmid SGC7901/EGFP-PRL-3 cells (high) by western blotting. (B) The expression of MMP-2 and MMP-9 were also analyzed by western blotting. (C) The relative expression of PRL-3 was quantified relative to β-actin. (D) The ratio of p-AKT/AKT was quantified relative to the control group. (E) The expression of MMP-2 was quantified relative to β-actin. (F) The expression of MMP-9 was quantified relative to β-actin. Each bar represents the mean ± standard deviation of 3 independent experiments. *P
    Mmp 9, supplied by Abcam, used in various techniques. Bioz Stars score: 98/100, based on 4223 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc antibodies mmp2
    PTH could regulate <t>MMP2</t> and MMP9 proteins expression by primary cilia.
    Antibodies Mmp2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti mmp9 antibody
    Downregulation of FOXK1 inhibits metastasis in vivo. ( A ) Lung tissues were obtained from nude mice injected with differently treated MGC803 cells (n=5). ( B ) The number of lung nodules per nude mouse was determined. ( C ) The lung weight of each nude mouse was calculated. ( D , E ) Representative images of H E staining of lung sections (scale bar, 100 μm and 50 μm) and IHC staining of FOXK1, LC3-II, P62, E-cadherin and <t>MMP9</t> levels in lung tissue (scale bar, 100 μm and 50 μm). ( F ) MGC803 cells transfected with LV-shFOXK1-1 and LV-shFOXK1-2 were orthotopically transplanted into the stomach walls of nude mice (n=5). ( G , H ) Weight of gastric wall tumors (g) and number of metastatic tumors ( H ) at 28 days post transplantation with MGC803 cells. ( I , J ) Representative images ( I ) and number ( J ) of diffuse liver metastases. * P
    Anti Mmp9 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 495 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore anti mmp 2
    Model of melatonin regulation of breast cancer cell invasion . Ligand-dependent activation of the membrane-associated G-protein-coupled receptor MT1 leads to coupling of G i2 protein to MT1 receptor. As a result, the Gα i2 subunit dissociates from the Gβγ subunits and inhibits the activity of adenylcyclase (AC), and this leads to a decrease in the intracellular level of cAMP and inhibition of protein kinase A (PKA) activity. The cAMP/PKA pathway cross-talks with the p38 pathway through PKA. In response to the reduced cAMP level, activity of p38 is suppressed, and this causes further downregulation of MMP-9 expression via repression of ETS1 transcriptional activity and, potentially, downregulation of <t>MMP-2</t> transcription. MMP, matrix metalloproteinase.
    Anti Mmp 2, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 234 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Abcam rabbit anti mmp 9
    Biochemical characterization of the osteoclast-likeness of the various MGCs. (A) Cell lysates were taken at indicated time points during the differentiation protocol and analyzed for <t>MMP-9</t> by Western blotting. 1* indicates lysates from RAW 264.7 cells prior to LPS/IFNγ or IL-4 addition. Numbers below MMP-9 blot represent densitometry readings for MMP-9 expression with 1.0 being day 2, RANKL-treated cells. (B) Cell lysates were taken from day 2 – day 4 during the differentiation protocol and analyzed for Cathepsin K expression. The last lane is the cell lysate from day 4 untreated RAW 264.7 cells. (C) Same as (B) but blots were analyzed for Arginase and iNOS (arrow). GAPDH was used as a loading control. Blots are representative of three independent experiments.
    Rabbit Anti Mmp 9, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 274 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore anti mmp 9 antibody
    <t>MMP-9</t> mediates Slug-dependent EMT in NRK52 cells downstream of TGF-β1. A–F: Immunofluorescence images showing E-cadherin ( A and B: Scale bar = 15 μm), α-SMA ( C and D) and β-catenin ( E and F ) of NRK52e cells
    Anti Mmp 9 Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 229 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc mmp9
    Co-treatment of Zyflamend and cisplatin decreased cell proliferation, increased apoptosis, and suppressed NFκB signaling in T24R xenografts. Notes: ( A ) Representative IHC staining of proliferative marker (Ki-67), apoptosis marker (cleaved caspase 3), RelA, and <t>MMP9</t> from the sections of vehicle, cisplatin, and/or Zyflamend treatment groups. Quantification of Ki-67-positive cancer cells ( B ) and cleaved caspase 3 ( C ) in the sections of T24R xenografts with different treatments. Data are presented as mean ± SE; * P
    Mmp9, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1555 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti mmp13 antibody
    Elimination of SnCs by ABT263 and alleviation inflammatory microenvironment in vivo. ( A ) immunohistochemical analysis for expression of p16 INK4a , HMGB1, COLII and ACAN of rat cartilage after 2 weeks intra-articular injection of ABT263. Result from ABT263 of 1.0 mM was presented. Scale bar: 100 μm. ( B ) mRNA level analysis using real-time qPCR for CDKN1A, CDKN2A, <t>MMP13,</t> ADAMTS5, IL1β, IL6, COLII and ACAN in the rat knee joint. Data are shown as mean ± standard deviation. N = 3 per group. +p
    Anti Mmp13 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 225 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore anti mt1 mmp
    Notch1 effects <t>MT1-MMP-dependent</t> melanoma cell growth. A , expression levels of MT1-MMP ( MT1 ) and Notch1 NIC in WM266-4 cells expressing shGFP or shMT1-MMP and transduced with active Notch1 ( NIC ). β- act , β-actin. B , relative growth at a
    Anti Mt1 Mmp, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 142 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam mmp 9 antibody
    Upregulation of <t>MMP-9</t> expression by CS. Notes: Immunohistochemical detection of MMP-9 (DAB, brown) in ( A ) CS-exposed mice and ( B ) air-exposed mice. Sections were counterstained with Mayer hematoxylin (blue). Scale bar=50 µm. ( C ) Number of MMP-9–positive cells per square millimeter, n=5. ( D ) MMP-9 mRNA level in the lung tissues, n=4. ( E ) Co-immunofluorescent staining for MMP-9 (AlexaFluor 594, red) and F4/80 (AlexaFluor 488, green) in the lungs of CS-exposed mice. Nuclei were stained with DAPI (blue). ( F and G ) Representative Western blot and relative quantification normalized to β-actin for MMP-9 in MH-S stimulated with CSE for 48 hours; n=3. Arrows indicate positive cells. Bars indicate the mean value, and error bars indicate SEM. * P
    Mmp 9 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 103 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    R&D Systems human mmp 1 antibody
    The schematic diagram of OA treatment by TGF-β3 and BMP7. ( A ) Osteoarthritic cartilage with a degenerative superficial layer and loss of chondrocytes shows higher catabolism, reflected by elevated <t>MMP-1/3/13</t> expression; ( B ) After treatment with the combination of TGF-β3 and BMP7, the superficial layer with chondrocytes with newly gained energy may exhibit higher anabolism, reflected by increased expression of COL2A1 and ACAN .
    Human Mmp 1 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 160 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems mouse mmp 9 antibody
    Adtrp deficiency leads to increased expression of matrix metallopeptidase‐9 (Mmp9) mRNA , protein, and activity levels in zebrafish embryos and newborn mice. A , Whole‐mount in situ hybridization for mmp9 (blue) in 72 hours postfertilization (hpf) control and adtrp zebrafish morphants. Blue arrow: specific signal accumulation. Black arrow: caudal vascular plexus ( CVP ). n, biological replicates. Bars, 100 μm. B , Immunofluorescence staining with <t>anti‐Mmp‐9</t> (Cy3, red) IgG on cross‐sectioned CVP (green) of 72‐hpf Tg(fli1: EGFP )y1 ( EGFP , fluorescent vascular reporter) control and adtrp morphants±human ADTRP mRNA . Bars, 100 μm. C , Total lysates of 72‐hpf zebrafish embryos incubated ±100 μmol/L of pan‐ MMP inhibitor GM 6001 for 24 hours and analyzed by 10% gelatin gel electrophoresis. Left: representative zymogram (grayscale inverted image). Histogram: semiquantitative densitometry and statistics (mean± SEM by t test). * P
    Mouse Mmp 9 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 280 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore anti mmp 1
    MMP-9 is involved in TGM2-mediated migration and invasion in the TRAIL-resistant cells. A , A549-TR cells were transfected with negative control siRNA or TGM2-siRNA and incubated for 48 h. Equal amounts of cell extracts were detected for TGM2, <t>MMP-1,</t> MMP-2,
    Anti Mmp 1, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 102 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effects of postischemia-reperfusion (I/R) treatment with TMZ on the expressions of MMPs and TIMPs. Western blot analysis (a) and densitometric analysis (b–e) of the Western blot for MMP-2, -9 and TIMP-1, -2 at 5 d, and 8 w after I/R injury, respectively. Post I/R treatment with TMZ reduced the degree of I/R-induced increases in MMP-2 and MMP-9 levels and further increased the levels of TIMP-1 and TIMP-2 at 5 d after renal I/R injury compared to the IRC group. At 8 w after I/R injury, MMP levels in the TMZ-treated groups were similar to those of the IRC groups, which were all significantly greater than those of the sham groups. TIMP levels in the TMZ-treated groups became also comparable to those of the IRC groups, which were significantly greater than those of the sham groups. TMZ = trimetazidine. Sham = rats not underwent I/R; IRC = rats underwent ischemia (45 min)-reperfusion (5 d or 8 w); TMZ S = rats treated with TMZ (3 mg/kg) upon reperfusion; TMZ D = rats treated with TMZ (3 mg/kg) once daily for 5 d or 8 w starting upon reperfusion. ∗ P

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Effects of Post Ischemia-Reperfusion Treatment with Trimetazidine on Renal Injury in Rats: Insights on Delayed Renal Fibrosis Progression

    doi: 10.1155/2018/1072805

    Figure Lengend Snippet: Effects of postischemia-reperfusion (I/R) treatment with TMZ on the expressions of MMPs and TIMPs. Western blot analysis (a) and densitometric analysis (b–e) of the Western blot for MMP-2, -9 and TIMP-1, -2 at 5 d, and 8 w after I/R injury, respectively. Post I/R treatment with TMZ reduced the degree of I/R-induced increases in MMP-2 and MMP-9 levels and further increased the levels of TIMP-1 and TIMP-2 at 5 d after renal I/R injury compared to the IRC group. At 8 w after I/R injury, MMP levels in the TMZ-treated groups were similar to those of the IRC groups, which were all significantly greater than those of the sham groups. TIMP levels in the TMZ-treated groups became also comparable to those of the IRC groups, which were significantly greater than those of the sham groups. TMZ = trimetazidine. Sham = rats not underwent I/R; IRC = rats underwent ischemia (45 min)-reperfusion (5 d or 8 w); TMZ S = rats treated with TMZ (3 mg/kg) upon reperfusion; TMZ D = rats treated with TMZ (3 mg/kg) once daily for 5 d or 8 w starting upon reperfusion. ∗ P

    Article Snippet: Proteins were separated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotted with anti-VEGF (Santa Cruz, CA), anti-matrix metalloproteinase- (MMP-) 2, anti-MMP-9, anti-tissue inhibitor of metalloproteinase- (TIMP-) 1, anti-TIMP-2 (Calbiochem, USA), anti-Bcl-2, anti-Bax, anti-HIF-1α , and anti-actin (all from Cell Signaling Technology, Beverly, MA).

    Techniques: Western Blot

    Mechanism of S. Typhimurium-induced intestinal damage during colitis. (A) Secretion of MMP-9 and MMP-2 in the colons of mock (Con)- or S. Typhimurium (SaI)-infected Lcn2 −/− mice was analyzed by gelatin zymography using gelatin-agarose-purified PBS extracts (6–8 mice per group). (B) Quantitation of gelatinolytic activities from the zymogram in panel A and from another representative zymogram from independent experiments. (C) MMP-9 protein levels in the colons of mice from the respective groups were assessed by immunoblotting purified PBS extracts under non-reducing conditions. Flow-through from the purification was used as a loading control. (D) Quantitation of changes in protein levels from the immunoblot in panel C and from another representative blot from independent experiments. (E) MMP-9 expression levels in the colons of mock- or S. Typhimurium-infected Lcn2 +/+ and Lcn2 −/− mice were measured by real-time PCR. Error bars = ± standard error of the mean. *p

    Journal: PLoS Pathogens

    Article Title: Absence of Intestinal PPAR? Aggravates Acute Infectious Colitis in Mice through a Lipocalin-2-Dependent Pathway

    doi: 10.1371/journal.ppat.1003887

    Figure Lengend Snippet: Mechanism of S. Typhimurium-induced intestinal damage during colitis. (A) Secretion of MMP-9 and MMP-2 in the colons of mock (Con)- or S. Typhimurium (SaI)-infected Lcn2 −/− mice was analyzed by gelatin zymography using gelatin-agarose-purified PBS extracts (6–8 mice per group). (B) Quantitation of gelatinolytic activities from the zymogram in panel A and from another representative zymogram from independent experiments. (C) MMP-9 protein levels in the colons of mice from the respective groups were assessed by immunoblotting purified PBS extracts under non-reducing conditions. Flow-through from the purification was used as a loading control. (D) Quantitation of changes in protein levels from the immunoblot in panel C and from another representative blot from independent experiments. (E) MMP-9 expression levels in the colons of mock- or S. Typhimurium-infected Lcn2 +/+ and Lcn2 −/− mice were measured by real-time PCR. Error bars = ± standard error of the mean. *p

    Article Snippet: Immunoblots were probed with anti-PPARγ (Cell Signaling), anti-MMP-9 (Abcam), anti-Lcn2 (Abcam), and anti-β-actin (Santa Cruz Biotechnology) antibodies.

    Techniques: Infection, Mouse Assay, Zymography, Purification, Quantitation Assay, Flow Cytometry, Expressing, Real-time Polymerase Chain Reaction

    Elevated levels of Lcn2 in the colonic milieu of S. Typhimurium-infected mice stabilize proMMP-9. (A) The activities of MMP-9 and MMP-2 in gelatin-agarose-purified PBS (secreted) extracts of mock (Con)- or S. Typhimurium (SaI)-infected PPARγVillinCre+ (Cre+) or PPARγVillinCre− (Cre−) mouse colonic tissues were analyzed by gelatin zymography (6–8 mice per group). (B) Quantitation of gelatinolytic activities from the zymogram in panel A and from two other representative zymograms from independent experiments. Expression levels of MMP-9 (C), MMP-2 (D), and TIMP-1 (E) in the colons of the respective groups were measured by real-time PCR. (F) Protein levels of MMP-9 and Lcn2 in the colons of the respective mouse groups were assessed by immunoblotting using purified PBS extracts under non-reducing conditions. Flow-through from gelatin-agarose purification was probed for β-actin as a loading control. (G) Quantitation of changes in protein levels from the immunoblot in panel F and from another representative blot from independent experiments. Error bars = ± standard error of the mean. *p

    Journal: PLoS Pathogens

    Article Title: Absence of Intestinal PPAR? Aggravates Acute Infectious Colitis in Mice through a Lipocalin-2-Dependent Pathway

    doi: 10.1371/journal.ppat.1003887

    Figure Lengend Snippet: Elevated levels of Lcn2 in the colonic milieu of S. Typhimurium-infected mice stabilize proMMP-9. (A) The activities of MMP-9 and MMP-2 in gelatin-agarose-purified PBS (secreted) extracts of mock (Con)- or S. Typhimurium (SaI)-infected PPARγVillinCre+ (Cre+) or PPARγVillinCre− (Cre−) mouse colonic tissues were analyzed by gelatin zymography (6–8 mice per group). (B) Quantitation of gelatinolytic activities from the zymogram in panel A and from two other representative zymograms from independent experiments. Expression levels of MMP-9 (C), MMP-2 (D), and TIMP-1 (E) in the colons of the respective groups were measured by real-time PCR. (F) Protein levels of MMP-9 and Lcn2 in the colons of the respective mouse groups were assessed by immunoblotting using purified PBS extracts under non-reducing conditions. Flow-through from gelatin-agarose purification was probed for β-actin as a loading control. (G) Quantitation of changes in protein levels from the immunoblot in panel F and from another representative blot from independent experiments. Error bars = ± standard error of the mean. *p

    Article Snippet: Immunoblots were probed with anti-PPARγ (Cell Signaling), anti-MMP-9 (Abcam), anti-Lcn2 (Abcam), and anti-β-actin (Santa Cruz Biotechnology) antibodies.

    Techniques: Infection, Mouse Assay, Purification, Zymography, Quantitation Assay, Expressing, Real-time Polymerase Chain Reaction, Flow Cytometry

    Effect of limb RIPc on transcription and expression of MMP-9 in ischemic brain. (A) Relative expression of MMP-9 mRNA (real-time quantitative reverse transcription-polymerase chain reaction). Results are expressed as fold change over the sham group. (B) Representative western blot images of MMP-9 protein in ischemic brain of different groups. (C) Protein expression levels of MMP-9. (D) Double immunofluorescence images of MMP-9 and GFAP in the MCAO group. Nuclei were labeled with DAPI (blue). MMP-9-positive cells were labeled green by Alexa Fluor 488, while GFAP-positive cells were labeled red by Alexa Fluor 594. White arrows indicate corresponding positive cells or co-localization of MMP-9 and GFAP. Scale bar: 20 μm. (E) Double immunofluorescence images of MMP-9 and NeuN in the MCAO group. Nuclei were labeled with DAPI (blue). MMP-9-positive cells were labeled green by Alexa Fluor 488, while NeuN-positive cells were labeled red by Alexa Fluor 594. White arrows indicate corresponding positive cells or co-localization of MMP-9 and NeuN. Scale bar: 20 μm. (F) Immunohistochemistry images of MMP-9. Black arrows indicate MMP-9-positive cells. Scale bar: 20 μm. (G) Number of MMP-9-positive cells around the infarct area. (A, C, G) Data are expressed as the mean ± SEM (one-way analysis of variance followed by Student’s t -test and post hoc Fisher’s tests). * P

    Journal: Neural Regeneration Research

    Article Title: Limb remote ischemic postconditioning protects integrity of the blood-brain barrier after stroke

    doi: 10.4103/1673-5374.237122

    Figure Lengend Snippet: Effect of limb RIPc on transcription and expression of MMP-9 in ischemic brain. (A) Relative expression of MMP-9 mRNA (real-time quantitative reverse transcription-polymerase chain reaction). Results are expressed as fold change over the sham group. (B) Representative western blot images of MMP-9 protein in ischemic brain of different groups. (C) Protein expression levels of MMP-9. (D) Double immunofluorescence images of MMP-9 and GFAP in the MCAO group. Nuclei were labeled with DAPI (blue). MMP-9-positive cells were labeled green by Alexa Fluor 488, while GFAP-positive cells were labeled red by Alexa Fluor 594. White arrows indicate corresponding positive cells or co-localization of MMP-9 and GFAP. Scale bar: 20 μm. (E) Double immunofluorescence images of MMP-9 and NeuN in the MCAO group. Nuclei were labeled with DAPI (blue). MMP-9-positive cells were labeled green by Alexa Fluor 488, while NeuN-positive cells were labeled red by Alexa Fluor 594. White arrows indicate corresponding positive cells or co-localization of MMP-9 and NeuN. Scale bar: 20 μm. (F) Immunohistochemistry images of MMP-9. Black arrows indicate MMP-9-positive cells. Scale bar: 20 μm. (G) Number of MMP-9-positive cells around the infarct area. (A, C, G) Data are expressed as the mean ± SEM (one-way analysis of variance followed by Student’s t -test and post hoc Fisher’s tests). * P

    Article Snippet: Afterwards, sections were incubated overnight at 4°C in mixtures of anti-MMP-9 antibody (rabbit, polyclonal, 1:50; Abcam)/anti-glial fibrillary acidic protein (GFAP) (mouse, monoclonal, 1:200; Proteintech), anti-claudin-5 antibody (rabbit, polyclonal, 1:100; Abcam)/anti-GFAP (mouse, monoclonal, 1:200; Proteintech), anti-MMP-9 antibody (rabbit, polyclonal, 1:50; Abcam)/anti-NeuN (mouse, monoclonal, 1:200; Abcam), anti-claudin-5 antibody (rabbit, polyclonal, 1:100; Abcam)/anti-NeuN (mouse, monoclonal, 1:200; Abcam), or anti-claudin-5 antibody (rabbit, polyclonal, 1:100; Abcam)/anti-MMP-9 (mouse, monoclonal, 1:50; Abcam).

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

    Effect of RPE treatment on levels of TGF-β1 and MMP-9 in BAL. TGF-βand MMP-9 in BALwere measured by ELISA. Data are means ± SD from triplicate experiments. ** P

    Journal: BMC Complementary and Alternative Medicine

    Article Title: Radix puerariae extracts ameliorate paraquat-induced pulmonary fibrosis by attenuating follistatin-like 1 and nuclear factor erythroid 2p45-related factor-2 signalling pathways through downregulation of miRNA-21 expression

    doi: 10.1186/s12906-016-0991-6

    Figure Lengend Snippet: Effect of RPE treatment on levels of TGF-β1 and MMP-9 in BAL. TGF-βand MMP-9 in BALwere measured by ELISA. Data are means ± SD from triplicate experiments. ** P

    Article Snippet: Membranes were blocked for 1 h at room temperature with 5 % bovine serum albumin (BSA) and then incubated overnight at 4 °C with the following primary antibodies: anti-MMP-9 (Santa Cruz Biotechnology, Dallas, TX, USA), anti-p-p38MAPK (Bio-Rad Laboratories Inc.), anti-NF-kB65 (DAKOCorp, Carpinteria, CA, USA), anti-HO-1 (Cell Signaling, Beverly, MA, USA), anti-Nrf2 (Cell Signaling), anti-FSTL1 (R & D systems, Minneapolis, MN, USA), anti-p-Smad2/3 (Serotec Ltd, Oxford, United Kingdom), anti-TGF-β1 (Sigma, St. Louis, MO, USA), anti-CTGF (Santa Cruz Biotechnology), anti-collagen III (Santa Cruz Biotechnology), anti-collagen-1 (Invitrogen), and anti-β-actin (Sigma-Aldrich), each diluted 1:1000 in Tris-buffered saline with Tween-20 (TBST). β-actin blotting served as the control to confirm protein loading.

    Techniques: Enzyme-linked Immunosorbent Assay

    Effects of RPE on miR-21, FSTL1, p-p38MAPK, NF-kB65, p-Smad2/3, and MMP-9 expression in lungs from mice treated with PQ. On day 14 after administration of RPE and PQ challenge, miR-21, FSTL1, p-p38MAPK, NF-kB65, p-Smad2/3 and MMP-9 expression were assessed by real-time PCR ( a ) and p-p38MAPK, NF-kB65, p-Smad2/3 and MMP-9 protein levels by western blotting ( b ) Groups are as defined in Methods. (a, control; b, model group; c, treatment group). Values are means ± SD as determined from three independent experiments. * P

    Journal: BMC Complementary and Alternative Medicine

    Article Title: Radix puerariae extracts ameliorate paraquat-induced pulmonary fibrosis by attenuating follistatin-like 1 and nuclear factor erythroid 2p45-related factor-2 signalling pathways through downregulation of miRNA-21 expression

    doi: 10.1186/s12906-016-0991-6

    Figure Lengend Snippet: Effects of RPE on miR-21, FSTL1, p-p38MAPK, NF-kB65, p-Smad2/3, and MMP-9 expression in lungs from mice treated with PQ. On day 14 after administration of RPE and PQ challenge, miR-21, FSTL1, p-p38MAPK, NF-kB65, p-Smad2/3 and MMP-9 expression were assessed by real-time PCR ( a ) and p-p38MAPK, NF-kB65, p-Smad2/3 and MMP-9 protein levels by western blotting ( b ) Groups are as defined in Methods. (a, control; b, model group; c, treatment group). Values are means ± SD as determined from three independent experiments. * P

    Article Snippet: Membranes were blocked for 1 h at room temperature with 5 % bovine serum albumin (BSA) and then incubated overnight at 4 °C with the following primary antibodies: anti-MMP-9 (Santa Cruz Biotechnology, Dallas, TX, USA), anti-p-p38MAPK (Bio-Rad Laboratories Inc.), anti-NF-kB65 (DAKOCorp, Carpinteria, CA, USA), anti-HO-1 (Cell Signaling, Beverly, MA, USA), anti-Nrf2 (Cell Signaling), anti-FSTL1 (R & D systems, Minneapolis, MN, USA), anti-p-Smad2/3 (Serotec Ltd, Oxford, United Kingdom), anti-TGF-β1 (Sigma, St. Louis, MO, USA), anti-CTGF (Santa Cruz Biotechnology), anti-collagen III (Santa Cruz Biotechnology), anti-collagen-1 (Invitrogen), and anti-β-actin (Sigma-Aldrich), each diluted 1:1000 in Tris-buffered saline with Tween-20 (TBST). β-actin blotting served as the control to confirm protein loading.

    Techniques: Expressing, Mouse Assay, Real-time Polymerase Chain Reaction, Western Blot

    CORM-2 reduces Ang II-induced MMP-9 expression via inhibition of the AT1R/IL-6 pathway in HASMCs. (A) Cells were treated with 10 μM Ang II for the indicated times, and then the production of IL-6 was measured. Cells were transfected with siRNA of scrambled or IL-6, and then treated with Ang II for 24 h. The concentration of MMP-9 was determined (B). The cell migration was determined by migration assay (C). (D) Cells were pretreated with HLN, NAC, APO, or CORM-2, and then treated with Ang II for 24 h. The production of IL-6 was measured. (E, F) Cells were transfected with siRNA of scrambled, AT1R, or AT2R, and then treated with Ang II for 24 h. The production of IL-6 and MMP-9 was measured. Data are expressed as mean±S.E.M. of three independent experiments. # P

    Journal: Redox Biology

    Article Title: CO-releasing molecules CORM2 attenuates angiotensin II-induced human aortic smooth muscle cell migration through inhibition of ROS/IL-6 generation and matrix metalloproteinases-9 expression

    doi: 10.1016/j.redox.2017.02.019

    Figure Lengend Snippet: CORM-2 reduces Ang II-induced MMP-9 expression via inhibition of the AT1R/IL-6 pathway in HASMCs. (A) Cells were treated with 10 μM Ang II for the indicated times, and then the production of IL-6 was measured. Cells were transfected with siRNA of scrambled or IL-6, and then treated with Ang II for 24 h. The concentration of MMP-9 was determined (B). The cell migration was determined by migration assay (C). (D) Cells were pretreated with HLN, NAC, APO, or CORM-2, and then treated with Ang II for 24 h. The production of IL-6 was measured. (E, F) Cells were transfected with siRNA of scrambled, AT1R, or AT2R, and then treated with Ang II for 24 h. The production of IL-6 and MMP-9 was measured. Data are expressed as mean±S.E.M. of three independent experiments. # P

    Article Snippet: Anti-GAPDH, anti-GαS, and anti-MMP-9 antibodies were purchased from Santa Cruz (Santa Cruz, CA).

    Techniques: Expressing, Inhibition, Transfection, Concentration Assay, Migration

    Schematic diagram illustrating the proposed signaling pathway involved in Ang II-induced AT1R/IL-6/MMP-9-dependent cell migration. In HASMCs, Ang II induces MMP-9 expression via the AT1R/NADPH oxidase/ROS/NF-κB/IL-6 pathway. Moreover, CORM-2 can inhibit cell migration via inhibition of the activation of AT1R/NADPH oxidase/ROS/NF-κB/IL-6/IL-6R/MMP-9.

    Journal: Redox Biology

    Article Title: CO-releasing molecules CORM2 attenuates angiotensin II-induced human aortic smooth muscle cell migration through inhibition of ROS/IL-6 generation and matrix metalloproteinases-9 expression

    doi: 10.1016/j.redox.2017.02.019

    Figure Lengend Snippet: Schematic diagram illustrating the proposed signaling pathway involved in Ang II-induced AT1R/IL-6/MMP-9-dependent cell migration. In HASMCs, Ang II induces MMP-9 expression via the AT1R/NADPH oxidase/ROS/NF-κB/IL-6 pathway. Moreover, CORM-2 can inhibit cell migration via inhibition of the activation of AT1R/NADPH oxidase/ROS/NF-κB/IL-6/IL-6R/MMP-9.

    Article Snippet: Anti-GAPDH, anti-GαS, and anti-MMP-9 antibodies were purchased from Santa Cruz (Santa Cruz, CA).

    Techniques: Migration, Expressing, Inhibition, Activation Assay

    CORM-2 reduces Ang II-induced NF-κB activation in HASMCs. Cells were transfected with siRNA of scrambled or p65, and then treated with Ang II for 24 h. The concentration of MMP-9 was determined (A). The cell migration was determined by migration assay (B). (C) Cells were pretreated with helenalin (HLN) or Bay11-7082 (Bay) for 1 h, and then treated with Ang II for 6 h. The mRNA levels and promoter activity of MMP-9 were determined by real-time PCR and promoter assay, respectively. (D) Cells were treated with 10 μM Ang II for the indicated times, and then the promoter activity of NF-κB was measured by promoter assay. (E) Cells were pretreated with CORM-2, NAC, APO, DPI, and HLN, and then treated with Ang II for 60 min. The promoter activity of NF-κB was measured by promoter assay. (F) Cells were pretreated with CORM-2, NAC, or HLN, and then treated with Ang II for the indicated times. The protein expression of phospho-p65 was determined. Data are expressed as mean±S.E.M. of three independent experiments. # P

    Journal: Redox Biology

    Article Title: CO-releasing molecules CORM2 attenuates angiotensin II-induced human aortic smooth muscle cell migration through inhibition of ROS/IL-6 generation and matrix metalloproteinases-9 expression

    doi: 10.1016/j.redox.2017.02.019

    Figure Lengend Snippet: CORM-2 reduces Ang II-induced NF-κB activation in HASMCs. Cells were transfected with siRNA of scrambled or p65, and then treated with Ang II for 24 h. The concentration of MMP-9 was determined (A). The cell migration was determined by migration assay (B). (C) Cells were pretreated with helenalin (HLN) or Bay11-7082 (Bay) for 1 h, and then treated with Ang II for 6 h. The mRNA levels and promoter activity of MMP-9 were determined by real-time PCR and promoter assay, respectively. (D) Cells were treated with 10 μM Ang II for the indicated times, and then the promoter activity of NF-κB was measured by promoter assay. (E) Cells were pretreated with CORM-2, NAC, APO, DPI, and HLN, and then treated with Ang II for 60 min. The promoter activity of NF-κB was measured by promoter assay. (F) Cells were pretreated with CORM-2, NAC, or HLN, and then treated with Ang II for the indicated times. The protein expression of phospho-p65 was determined. Data are expressed as mean±S.E.M. of three independent experiments. # P

    Article Snippet: Anti-GAPDH, anti-GαS, and anti-MMP-9 antibodies were purchased from Santa Cruz (Santa Cruz, CA).

    Techniques: Activation Assay, Transfection, Concentration Assay, Migration, Activity Assay, Real-time Polymerase Chain Reaction, Promoter Assay, Expressing

    Ang II induces MMP-9-dependent cell migration. (A) Cells were incubated with Ang II for the indicated times, and then the cell viability was determined. (B) HASMCs were treated with Ang II for 24 h, and then the mRNA levels of AT1R and AT2R were determined by RT-PCR. (C) Cells were incubated with Ang II for the indicated times, and then the protein expression of MMP-9 was determined by Western blot. (D, E) Cells were incubated with Ang II (10 μM) for the indicated times, and then the mRNA levels and promoter activity of MMP-9 were determined by real-time PCR and promoter assay, respectively. (F) Cells were incubated with Ang II (10 μM) for the indicated times or transfected with siRNA of scrambled or MMP-9, and then treated with Ang II (10 μM) for 24 h. The cell migration was determined by migration assay. Data are expressed as mean±S.E.M. of three independent experiments. *P

    Journal: Redox Biology

    Article Title: CO-releasing molecules CORM2 attenuates angiotensin II-induced human aortic smooth muscle cell migration through inhibition of ROS/IL-6 generation and matrix metalloproteinases-9 expression

    doi: 10.1016/j.redox.2017.02.019

    Figure Lengend Snippet: Ang II induces MMP-9-dependent cell migration. (A) Cells were incubated with Ang II for the indicated times, and then the cell viability was determined. (B) HASMCs were treated with Ang II for 24 h, and then the mRNA levels of AT1R and AT2R were determined by RT-PCR. (C) Cells were incubated with Ang II for the indicated times, and then the protein expression of MMP-9 was determined by Western blot. (D, E) Cells were incubated with Ang II (10 μM) for the indicated times, and then the mRNA levels and promoter activity of MMP-9 were determined by real-time PCR and promoter assay, respectively. (F) Cells were incubated with Ang II (10 μM) for the indicated times or transfected with siRNA of scrambled or MMP-9, and then treated with Ang II (10 μM) for 24 h. The cell migration was determined by migration assay. Data are expressed as mean±S.E.M. of three independent experiments. *P

    Article Snippet: Anti-GAPDH, anti-GαS, and anti-MMP-9 antibodies were purchased from Santa Cruz (Santa Cruz, CA).

    Techniques: Migration, Incubation, Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, Activity Assay, Real-time Polymerase Chain Reaction, Promoter Assay, Transfection

    Ang II induces MMP-9 expression via NADPH oxidase/ROS in HASMCs. (A) Cells were transfected with siRNA of scrambled, Nox2, or Nox4, and then treated with Ang II for 24 h. The conditioned media were subjected to determine MMP-9 expression by gelatin zymography. (B) Cells were transfected with siRNA of scrambled, p47 phox , Nox2, or Nox4, and then treated with Ang II for 24 h. The concentration of MMP-9 was determined. (C) Cells were transfected with siRNA of scrambled, p47 phox , Nox2, or Nox4, and then treated with Ang II for 24 h. The cell migration was determined by migration assay. (D) Cells were pretreated with NAC, DPI, or APO for 1 h, and then treated with Ang II for 6 h. The mRNA levels and promoter activity of MMP-9 were determined by real-time PCR and promoter assay, respectively. Data are expressed as mean±S.E.M. of three independent experiments. *P

    Journal: Redox Biology

    Article Title: CO-releasing molecules CORM2 attenuates angiotensin II-induced human aortic smooth muscle cell migration through inhibition of ROS/IL-6 generation and matrix metalloproteinases-9 expression

    doi: 10.1016/j.redox.2017.02.019

    Figure Lengend Snippet: Ang II induces MMP-9 expression via NADPH oxidase/ROS in HASMCs. (A) Cells were transfected with siRNA of scrambled, Nox2, or Nox4, and then treated with Ang II for 24 h. The conditioned media were subjected to determine MMP-9 expression by gelatin zymography. (B) Cells were transfected with siRNA of scrambled, p47 phox , Nox2, or Nox4, and then treated with Ang II for 24 h. The concentration of MMP-9 was determined. (C) Cells were transfected with siRNA of scrambled, p47 phox , Nox2, or Nox4, and then treated with Ang II for 24 h. The cell migration was determined by migration assay. (D) Cells were pretreated with NAC, DPI, or APO for 1 h, and then treated with Ang II for 6 h. The mRNA levels and promoter activity of MMP-9 were determined by real-time PCR and promoter assay, respectively. Data are expressed as mean±S.E.M. of three independent experiments. *P

    Article Snippet: Anti-GAPDH, anti-GαS, and anti-MMP-9 antibodies were purchased from Santa Cruz (Santa Cruz, CA).

    Techniques: Expressing, Transfection, Zymography, Concentration Assay, Migration, Activity Assay, Real-time Polymerase Chain Reaction, Promoter Assay

    CORM-2 inhibits Ang II-induced MMP-9 expression and cell migration. (A) Cells were incubated with CORM-2 for the indicated times, and then the cell viability was determined. (B) Cells were pretreated with CORM-2 for 2 h in the presence or absence of hemoglobin (Hb), and then treated with Ang II for 24 h. The concentration of MMP-9 was determined. (C) Cells were pretreated with CORM-2 for 2 h, and then treated with Ang II for 24 h. The conditioned media were subjected to determine MMP-9 expression by gelatin zymography. (D) Cells were pretreated with CORM-2 for 2 h in the presence or absence of Hb, and then treated with Ang II for 24 h. The cell migration was determined by migration assay. (E) Cells were pretreated with CORM-2 for 2 h in the presence or absence of Hb, and then treated with Ang II for 6 h. The mRNA levels and promoter activity of MMP-9 were determined by real-time PCR and promoter assay, respectively. Data are expressed as mean±S.E.M. of three independent experiments. # P

    Journal: Redox Biology

    Article Title: CO-releasing molecules CORM2 attenuates angiotensin II-induced human aortic smooth muscle cell migration through inhibition of ROS/IL-6 generation and matrix metalloproteinases-9 expression

    doi: 10.1016/j.redox.2017.02.019

    Figure Lengend Snippet: CORM-2 inhibits Ang II-induced MMP-9 expression and cell migration. (A) Cells were incubated with CORM-2 for the indicated times, and then the cell viability was determined. (B) Cells were pretreated with CORM-2 for 2 h in the presence or absence of hemoglobin (Hb), and then treated with Ang II for 24 h. The concentration of MMP-9 was determined. (C) Cells were pretreated with CORM-2 for 2 h, and then treated with Ang II for 24 h. The conditioned media were subjected to determine MMP-9 expression by gelatin zymography. (D) Cells were pretreated with CORM-2 for 2 h in the presence or absence of Hb, and then treated with Ang II for 24 h. The cell migration was determined by migration assay. (E) Cells were pretreated with CORM-2 for 2 h in the presence or absence of Hb, and then treated with Ang II for 6 h. The mRNA levels and promoter activity of MMP-9 were determined by real-time PCR and promoter assay, respectively. Data are expressed as mean±S.E.M. of three independent experiments. # P

    Article Snippet: Anti-GAPDH, anti-GαS, and anti-MMP-9 antibodies were purchased from Santa Cruz (Santa Cruz, CA).

    Techniques: Expressing, Migration, Incubation, Concentration Assay, Zymography, Activity Assay, Real-time Polymerase Chain Reaction, Promoter Assay

    Effects of the ERK inhibitor (U0126) and baicalein on cell invasion and MMP-2, MMP-9, u-PA, TIMP-1 and TIMP-2 expression in MHCC97H cells. (A) Cells were pretreated with U0126 (10 μM) for 30 min and then incubated in the presence or absence of baicalein (10 μM) for 24 h. Cellular invasiveness was measured using the Boyden chamber invasion assay. (B) The percent invasion rate was expressed as a percentage of control. (C, D) MHCC97H cells were treated and then subjected to western blotting to analyze the protein levels of MMP-2, MMP-9, u-PA, TIMP-1 and TIMP-2. Values represent the means ± SD of three independent experiments performed in triplicate. * p

    Journal: PLoS ONE

    Article Title: Baicalein Inhibits the Invasion and Metastatic Capabilities of Hepatocellular Carcinoma Cells via Down-Regulation of the ERK Pathway

    doi: 10.1371/journal.pone.0072927

    Figure Lengend Snippet: Effects of the ERK inhibitor (U0126) and baicalein on cell invasion and MMP-2, MMP-9, u-PA, TIMP-1 and TIMP-2 expression in MHCC97H cells. (A) Cells were pretreated with U0126 (10 μM) for 30 min and then incubated in the presence or absence of baicalein (10 μM) for 24 h. Cellular invasiveness was measured using the Boyden chamber invasion assay. (B) The percent invasion rate was expressed as a percentage of control. (C, D) MHCC97H cells were treated and then subjected to western blotting to analyze the protein levels of MMP-2, MMP-9, u-PA, TIMP-1 and TIMP-2. Values represent the means ± SD of three independent experiments performed in triplicate. * p

    Article Snippet: Anti-Phospho-MEK1 (Thr386) (p-MEK1) and anti-MEK1 were purchased from Millipore Co. Anti-Phospho (Thr202/Tyr204) ERK1/2 (p-ERK1/2), anti-ERK1/2, anti-MMP-2, anti-MMP-9, anti-TIMP-1 and anti-TIMP-2 antibodies were purchased from Cell Signaling.

    Techniques: Expressing, Incubation, Invasion Assay, Western Blot

    Baicalein suppresses the expression and activity of MMP-2, MMP-9 and u-PA and promotes the expression of TIMP-1 and TIMP-2 in MHCC97H cells. (A) The effects of baicalein on the expression levels of MMP-2, MMP-9 and u-PA mRNA were assessed by qRT-PCR. (B) MHCC97H cells were treated with baicalein (0, 10, 20 and 30 µM) for 24 h and then subjected to western blotting to analyze the protein levels of MMP-2, MMP-9 and u-PA. (C) Quantification of the protein levels of MMP-2, MMP-9 and u-PA. (D) Effects of baicalein on the activities of MMP-2, MMP-9 and u-PA. (E) Quantification of the activities of MMP-2, MMP-9 and u-PA. (F) MHCC97H cells were treated with baicalein (0, 10, 20 and 30 µM) for 24 h and then subjected to western blotting to analyze the protein levels of TIMP-1 and TIMP-2. (G) Quantification of the protein levels of TIMP-1 and TIMP-2. Values represent the means ± SD of three independent experiments performed in triplicate. * p

    Journal: PLoS ONE

    Article Title: Baicalein Inhibits the Invasion and Metastatic Capabilities of Hepatocellular Carcinoma Cells via Down-Regulation of the ERK Pathway

    doi: 10.1371/journal.pone.0072927

    Figure Lengend Snippet: Baicalein suppresses the expression and activity of MMP-2, MMP-9 and u-PA and promotes the expression of TIMP-1 and TIMP-2 in MHCC97H cells. (A) The effects of baicalein on the expression levels of MMP-2, MMP-9 and u-PA mRNA were assessed by qRT-PCR. (B) MHCC97H cells were treated with baicalein (0, 10, 20 and 30 µM) for 24 h and then subjected to western blotting to analyze the protein levels of MMP-2, MMP-9 and u-PA. (C) Quantification of the protein levels of MMP-2, MMP-9 and u-PA. (D) Effects of baicalein on the activities of MMP-2, MMP-9 and u-PA. (E) Quantification of the activities of MMP-2, MMP-9 and u-PA. (F) MHCC97H cells were treated with baicalein (0, 10, 20 and 30 µM) for 24 h and then subjected to western blotting to analyze the protein levels of TIMP-1 and TIMP-2. (G) Quantification of the protein levels of TIMP-1 and TIMP-2. Values represent the means ± SD of three independent experiments performed in triplicate. * p

    Article Snippet: Anti-Phospho-MEK1 (Thr386) (p-MEK1) and anti-MEK1 were purchased from Millipore Co. Anti-Phospho (Thr202/Tyr204) ERK1/2 (p-ERK1/2), anti-ERK1/2, anti-MMP-2, anti-MMP-9, anti-TIMP-1 and anti-TIMP-2 antibodies were purchased from Cell Signaling.

    Techniques: Expressing, Activity Assay, Quantitative RT-PCR, Western Blot

    Down-regulation of S100A4 inhibited cell proliferation of A7r5 and MMP2/MMP9 expression in vitro . A, The results of cell proliferation ability of A7r5 cells after transfection with S100A4 siRNA at different time intervals (0, 24, 48, 72 h). *Represents P

    Journal: PLoS ONE

    Article Title: Spatiotemporal Expression of Matrix Metalloproteinases (MMPs) is Regulated by the Ca2+-Signal Transducer S100A4 in the Pathogenesis of Thoracic Aortic Aneurysm

    doi: 10.1371/journal.pone.0070057

    Figure Lengend Snippet: Down-regulation of S100A4 inhibited cell proliferation of A7r5 and MMP2/MMP9 expression in vitro . A, The results of cell proliferation ability of A7r5 cells after transfection with S100A4 siRNA at different time intervals (0, 24, 48, 72 h). *Represents P

    Article Snippet: Primary antibodies include anti-MMP2, anti-MMP9, anti-GAPDH (all from Cell Signaling Technology).

    Techniques: Expressing, In Vitro, Transfection

    Curcumin retards activation of β-catenin/Slug pathway in breast cancer stem cells, thereby restoring E-cadherin. (A) β-catenin-associated E-cadherin was assayed by co-immunoprecipitation from cell lysates of MCF-7-derived 2° mammospheres with or without curcumin treatment using specific antibodies (left panel) or with normal human immunoglobulin G (IgG) as a negative control (right panel). To ensure comparable protein loading, 20% of supernatant from immunoprecipitation (IP) sample was subjected to determination of α-actin by Western blotting. (B) Western blotting was conducted to study the levels of total β-catenin and nuclear β-catenin in 2° mammospheres in presence or absence of curcumin exposure. (C) The relative nuclear expression of β-catenin in 2° mammospheres with or without curcumin treatment was visualized by immunofluorescence. (D, E) Under similar conditions, Western blotting was performed to study the expression levels of β-catenin target genes Cyclin-D1, c-Myc, Slug, Snail, Vimentin, MMP-2 and MMP-9 in curcumin treated/untreated 2° mammospheres. (F) Protein expression of E-cadherin in 2° mammospheres with or without curcumin or Slug-cDNA (or both) were determined by Western blotting (left panel). The efficiency of transfection was determined by Western blot analysis (right panel). (G) Graphical quantifications of cell adhesion, spreading, three-dimensional (3D) invasion, and migration assays of MCF-7-derived 2° mammospheres with or without curcumin and Slug-cDNA treatment/transfection. (H) Transwell migration assay was performed under similar experimental conditions in T47D-derived 2° mammospheres. α-Actin/histone H1/glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal loading control. Data are presented as mean ± standard error of mean or representative of three independent experiments. Cont, control; Cur, curcumin.

    Journal: Stem Cell Research & Therapy

    Article Title: Curcumin inhibits breast cancer stem cell migration by amplifying the E-cadherin/β-catenin negative feedback loop

    doi: 10.1186/scrt506

    Figure Lengend Snippet: Curcumin retards activation of β-catenin/Slug pathway in breast cancer stem cells, thereby restoring E-cadherin. (A) β-catenin-associated E-cadherin was assayed by co-immunoprecipitation from cell lysates of MCF-7-derived 2° mammospheres with or without curcumin treatment using specific antibodies (left panel) or with normal human immunoglobulin G (IgG) as a negative control (right panel). To ensure comparable protein loading, 20% of supernatant from immunoprecipitation (IP) sample was subjected to determination of α-actin by Western blotting. (B) Western blotting was conducted to study the levels of total β-catenin and nuclear β-catenin in 2° mammospheres in presence or absence of curcumin exposure. (C) The relative nuclear expression of β-catenin in 2° mammospheres with or without curcumin treatment was visualized by immunofluorescence. (D, E) Under similar conditions, Western blotting was performed to study the expression levels of β-catenin target genes Cyclin-D1, c-Myc, Slug, Snail, Vimentin, MMP-2 and MMP-9 in curcumin treated/untreated 2° mammospheres. (F) Protein expression of E-cadherin in 2° mammospheres with or without curcumin or Slug-cDNA (or both) were determined by Western blotting (left panel). The efficiency of transfection was determined by Western blot analysis (right panel). (G) Graphical quantifications of cell adhesion, spreading, three-dimensional (3D) invasion, and migration assays of MCF-7-derived 2° mammospheres with or without curcumin and Slug-cDNA treatment/transfection. (H) Transwell migration assay was performed under similar experimental conditions in T47D-derived 2° mammospheres. α-Actin/histone H1/glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal loading control. Data are presented as mean ± standard error of mean or representative of three independent experiments. Cont, control; Cur, curcumin.

    Article Snippet: For direct Western blot analysis, the cell lysates or the particular fractions were separated by SDS-PAGE, transferred to polyvinylidene difluoride membrane (Millipore, Darmstadt, Germany), and probed with specific antibodies like anti-E-cadherin, anti-β-catenin, anti-histone H1, anti-cyclin-D1, anti-c-myc, anti-slug, anti-vimentin, anti-MMP-2, anti-MMP-9, anti-twist, anti-Snail, and anti-α-Actin (Santa Cruz Biotechnology, Inc.).

    Techniques: Activation Assay, Immunoprecipitation, Derivative Assay, Negative Control, Western Blot, Expressing, Immunofluorescence, Transfection, Migration, Transwell Migration Assay

    TLR2 blockade inhibits vascular inflammation in atherosclerotic arteries. MMP-2 expression (A and B), NF-κB p65 phosphorylation (C), Stat3 phosphorylation (D) and the expression of the cytokines IL-6, TNF-α, and IL-10 (E) were reduced

    Journal: Acta Pharmacologica Sinica

    Article Title: Blocking TLR2 activity diminishes and stabilizes advanced atherosclerotic lesions in apolipoprotein E-deficient mice

    doi: 10.1038/aps.2013.75

    Figure Lengend Snippet: TLR2 blockade inhibits vascular inflammation in atherosclerotic arteries. MMP-2 expression (A and B), NF-κB p65 phosphorylation (C), Stat3 phosphorylation (D) and the expression of the cytokines IL-6, TNF-α, and IL-10 (E) were reduced

    Article Snippet: After tissue homogenization, the extracted proteins were separated using 12% SDS-PAGE and subjected to immunoblotting with specific an tibodies, including anti-MMP-2 (Abcam, Cambridge, UK), anti-phospho-NF-κB p65, anti-NF-κB p65, anti-cleaved caspase-3, anti-CHOP (Cell Signaling Technology, Danvers, MA, USA), anti-phospho-Stat3, anti-Stat3, anti-interleukin (IL)-6, anti-IL-10 or anti-tumor necrosis factor (TNF)-α (Santa Cruz Biotechnology, CA, USA).

    Techniques: Expressing

    UPA treatment upregulates MMP-2 expression. (A) mRNA expression of MMP-2 in leiomyoma cells treated with UPA. (B) Protein expression levels of MMP-9 and MMP-2 in leiomyoma cells treated with UPA. (C) Representative immunohistochemistry staining of MMP-9 and MMP-2 in uterine samples of patients (n=4) with no UPA treatment (CTL) and with UPA treatment (UPA) prior to hysterectomy (n=3). Data are presented as mean ± standard deviation. * P

    Journal: International Journal of Molecular Medicine

    Article Title: Ulipristal acetate induces cell cycle delay and remodeling of extracellular matrix

    doi: 10.3892/ijmm.2018.3779

    Figure Lengend Snippet: UPA treatment upregulates MMP-2 expression. (A) mRNA expression of MMP-2 in leiomyoma cells treated with UPA. (B) Protein expression levels of MMP-9 and MMP-2 in leiomyoma cells treated with UPA. (C) Representative immunohistochemistry staining of MMP-9 and MMP-2 in uterine samples of patients (n=4) with no UPA treatment (CTL) and with UPA treatment (UPA) prior to hysterectomy (n=3). Data are presented as mean ± standard deviation. * P

    Article Snippet: Membranes were blocked with 5% skim milk in TBS containing 0.1% Tween-20 for 1 h. Antibodies against MMP-9 (cat. no. ab38898), MMP-2 (cat. no. ab37150), CDK2 (cat. no. ab32147), cyclin E1 (cat. no. ab3927) were obtained from Abcam (Cambridge, MA, USA), for p21 (cat. no. 2947) and p27 (cat. no. 2552) from Cell Signaling Technology, Inc. (Danvers, MA, USA) and for β-actin (cat. no. A5316) from Sigma-Aldrich; Merck KGaA.

    Techniques: Expressing, Immunohistochemistry, Staining, CTL Assay, Standard Deviation

    PCP increases MMP and HSP-27 protein expression in pulmonary tissue. Immunohistochemical analysis for MMP2, MMP8, MMP9, MMP12 and HSP-27 expression in lung tissues of control and PCP groups. Positive protein staining appears brown, while nuclear staining appears blue. Magnification, x400. PCP, Pneumocystis pneumonia; MMP, matrix metalloprotease; HSP, heat shock protein.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Role of Pneumocystis jirovecii infection in chronic obstructive pulmonary disease progression in an immunosuppressed rat Pneumocystis pneumonia model

    doi: 10.3892/etm.2020.8545

    Figure Lengend Snippet: PCP increases MMP and HSP-27 protein expression in pulmonary tissue. Immunohistochemical analysis for MMP2, MMP8, MMP9, MMP12 and HSP-27 expression in lung tissues of control and PCP groups. Positive protein staining appears brown, while nuclear staining appears blue. Magnification, x400. PCP, Pneumocystis pneumonia; MMP, matrix metalloprotease; HSP, heat shock protein.

    Article Snippet: The membranes were incubated with the following primary antibodies diluted in 5% BSA TBST: Anti-MMP-2 (1:2,000; cat. no. ab37150; Abcam), anti-MMP-8 (1:2,000; cat. no. ab81286; Abcam), anti-MMP-9 (1:2,000; cat. no. ab76003; Abcam), anti-MMP-12 (1:2,000; cat. no. ab52897; Abcam) and anti-HSP-27 (1:1,000; cat. no. ab5579; Abcam) then incubated overnight at 4˚C.

    Techniques: Expressing, Immunohistochemistry, Staining

    PCP significantly increases MMP and HSP-27 levels in pulmonary tissue. IOD values from the immunohistochemistry staining for MMP2, MMP8, MMP9, MMP12 and HSP-27. * * P

    Journal: Experimental and Therapeutic Medicine

    Article Title: Role of Pneumocystis jirovecii infection in chronic obstructive pulmonary disease progression in an immunosuppressed rat Pneumocystis pneumonia model

    doi: 10.3892/etm.2020.8545

    Figure Lengend Snippet: PCP significantly increases MMP and HSP-27 levels in pulmonary tissue. IOD values from the immunohistochemistry staining for MMP2, MMP8, MMP9, MMP12 and HSP-27. * * P

    Article Snippet: The membranes were incubated with the following primary antibodies diluted in 5% BSA TBST: Anti-MMP-2 (1:2,000; cat. no. ab37150; Abcam), anti-MMP-8 (1:2,000; cat. no. ab81286; Abcam), anti-MMP-9 (1:2,000; cat. no. ab76003; Abcam), anti-MMP-12 (1:2,000; cat. no. ab52897; Abcam) and anti-HSP-27 (1:1,000; cat. no. ab5579; Abcam) then incubated overnight at 4˚C.

    Techniques: Immunohistochemistry, Staining

    p-AKT, MMP-2 and MMP-9 are involved in PRL-3-mediated invasion and migration of gastric cancer cells. (A) Levels of PRL-3, AKT and p-AKT were analyzed in SGC7901 cells (control), SGC7901 cells transfected with empty vector, and plasmid SGC7901/EGFP-PRL-3 cells (high) by western blotting. (B) The expression of MMP-2 and MMP-9 were also analyzed by western blotting. (C) The relative expression of PRL-3 was quantified relative to β-actin. (D) The ratio of p-AKT/AKT was quantified relative to the control group. (E) The expression of MMP-2 was quantified relative to β-actin. (F) The expression of MMP-9 was quantified relative to β-actin. Each bar represents the mean ± standard deviation of 3 independent experiments. *P

    Journal: Oncology Letters

    Article Title: PRL-3 promotes gastric cancer peritoneal metastasis via the PI3K/AKT signaling pathway in vitro and in vivo

    doi: 10.3892/ol.2018.8467

    Figure Lengend Snippet: p-AKT, MMP-2 and MMP-9 are involved in PRL-3-mediated invasion and migration of gastric cancer cells. (A) Levels of PRL-3, AKT and p-AKT were analyzed in SGC7901 cells (control), SGC7901 cells transfected with empty vector, and plasmid SGC7901/EGFP-PRL-3 cells (high) by western blotting. (B) The expression of MMP-2 and MMP-9 were also analyzed by western blotting. (C) The relative expression of PRL-3 was quantified relative to β-actin. (D) The ratio of p-AKT/AKT was quantified relative to the control group. (E) The expression of MMP-2 was quantified relative to β-actin. (F) The expression of MMP-9 was quantified relative to β-actin. Each bar represents the mean ± standard deviation of 3 independent experiments. *P

    Article Snippet: Antibodies against PRL-3 (ab50276; 1:400), AKT (ab8805; 1:500), p-AKT S473 (ab81283; 1:1,000), MMP-2 (ab97779; 1:1,000) and MMP-9 (ab137867; 1:1,000) were purchased from Abcam (Cambridge, UK).

    Techniques: Migration, Transfection, Plasmid Preparation, Western Blot, Expressing, Standard Deviation

    PTH could regulate MMP2 and MMP9 proteins expression by primary cilia.

    Journal: International Journal of Molecular Sciences

    Article Title: Evaluating the Role of PTH in Promotion of Chondrosarcoma Cell Proliferation and Invasion by Inhibiting Primary Cilia Expression

    doi: 10.3390/ijms151119816

    Figure Lengend Snippet: PTH could regulate MMP2 and MMP9 proteins expression by primary cilia.

    Article Snippet: We incubated these membranes overnight with primary antibodies-MMP2 (1:1000 dilution, Cell Signaling Technology, Danvers, MA, USA); MMP9 (1:1000 dilution, Cell Signaling Technology, Danvers, MA, USA); IFT88 (1:500 dilution, ABGENT, San Diego, CA, USA); PTHrP (1:100 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA); β-actin (1:400 dilution, Boster, Wuhan, China)-following up with secondary antibody horse-radish peroxidase-labeled goat anti-rabbit and goat anti-mouse (1:5000 dilution, Boster, Wuhan, China) IgG for one hour.

    Techniques: Expressing

    Effects of the ERK inhibitor (U0126) and baicalein on cell invasion and MMP-2, MMP-9, u-PA, TIMP-1 and TIMP-2 expression in MHCC97H cells. (A) Cells were pretreated with U0126 (10 μM) for 30 min and then incubated in the presence or absence of baicalein (10 μM) for 24 h. Cellular invasiveness was measured using the Boyden chamber invasion assay. (B) The percent invasion rate was expressed as a percentage of control. (C, D) MHCC97H cells were treated and then subjected to western blotting to analyze the protein levels of MMP-2, MMP-9, u-PA, TIMP-1 and TIMP-2. Values represent the means ± SD of three independent experiments performed in triplicate. * p

    Journal: PLoS ONE

    Article Title: Baicalein Inhibits the Invasion and Metastatic Capabilities of Hepatocellular Carcinoma Cells via Down-Regulation of the ERK Pathway

    doi: 10.1371/journal.pone.0072927

    Figure Lengend Snippet: Effects of the ERK inhibitor (U0126) and baicalein on cell invasion and MMP-2, MMP-9, u-PA, TIMP-1 and TIMP-2 expression in MHCC97H cells. (A) Cells were pretreated with U0126 (10 μM) for 30 min and then incubated in the presence or absence of baicalein (10 μM) for 24 h. Cellular invasiveness was measured using the Boyden chamber invasion assay. (B) The percent invasion rate was expressed as a percentage of control. (C, D) MHCC97H cells were treated and then subjected to western blotting to analyze the protein levels of MMP-2, MMP-9, u-PA, TIMP-1 and TIMP-2. Values represent the means ± SD of three independent experiments performed in triplicate. * p

    Article Snippet: Anti-Phospho-MEK1 (Thr386) (p-MEK1) and anti-MEK1 were purchased from Millipore Co. Anti-Phospho (Thr202/Tyr204) ERK1/2 (p-ERK1/2), anti-ERK1/2, anti-MMP-2, anti-MMP-9, anti-TIMP-1 and anti-TIMP-2 antibodies were purchased from Cell Signaling.

    Techniques: Expressing, Incubation, Invasion Assay, Western Blot

    Baicalein suppresses the expression and activity of MMP-2, MMP-9 and u-PA and promotes the expression of TIMP-1 and TIMP-2 in MHCC97H cells. (A) The effects of baicalein on the expression levels of MMP-2, MMP-9 and u-PA mRNA were assessed by qRT-PCR. (B) MHCC97H cells were treated with baicalein (0, 10, 20 and 30 µM) for 24 h and then subjected to western blotting to analyze the protein levels of MMP-2, MMP-9 and u-PA. (C) Quantification of the protein levels of MMP-2, MMP-9 and u-PA. (D) Effects of baicalein on the activities of MMP-2, MMP-9 and u-PA. (E) Quantification of the activities of MMP-2, MMP-9 and u-PA. (F) MHCC97H cells were treated with baicalein (0, 10, 20 and 30 µM) for 24 h and then subjected to western blotting to analyze the protein levels of TIMP-1 and TIMP-2. (G) Quantification of the protein levels of TIMP-1 and TIMP-2. Values represent the means ± SD of three independent experiments performed in triplicate. * p

    Journal: PLoS ONE

    Article Title: Baicalein Inhibits the Invasion and Metastatic Capabilities of Hepatocellular Carcinoma Cells via Down-Regulation of the ERK Pathway

    doi: 10.1371/journal.pone.0072927

    Figure Lengend Snippet: Baicalein suppresses the expression and activity of MMP-2, MMP-9 and u-PA and promotes the expression of TIMP-1 and TIMP-2 in MHCC97H cells. (A) The effects of baicalein on the expression levels of MMP-2, MMP-9 and u-PA mRNA were assessed by qRT-PCR. (B) MHCC97H cells were treated with baicalein (0, 10, 20 and 30 µM) for 24 h and then subjected to western blotting to analyze the protein levels of MMP-2, MMP-9 and u-PA. (C) Quantification of the protein levels of MMP-2, MMP-9 and u-PA. (D) Effects of baicalein on the activities of MMP-2, MMP-9 and u-PA. (E) Quantification of the activities of MMP-2, MMP-9 and u-PA. (F) MHCC97H cells were treated with baicalein (0, 10, 20 and 30 µM) for 24 h and then subjected to western blotting to analyze the protein levels of TIMP-1 and TIMP-2. (G) Quantification of the protein levels of TIMP-1 and TIMP-2. Values represent the means ± SD of three independent experiments performed in triplicate. * p

    Article Snippet: Anti-Phospho-MEK1 (Thr386) (p-MEK1) and anti-MEK1 were purchased from Millipore Co. Anti-Phospho (Thr202/Tyr204) ERK1/2 (p-ERK1/2), anti-ERK1/2, anti-MMP-2, anti-MMP-9, anti-TIMP-1 and anti-TIMP-2 antibodies were purchased from Cell Signaling.

    Techniques: Expressing, Activity Assay, Quantitative RT-PCR, Western Blot

    Downregulation of FOXK1 inhibits metastasis in vivo. ( A ) Lung tissues were obtained from nude mice injected with differently treated MGC803 cells (n=5). ( B ) The number of lung nodules per nude mouse was determined. ( C ) The lung weight of each nude mouse was calculated. ( D , E ) Representative images of H E staining of lung sections (scale bar, 100 μm and 50 μm) and IHC staining of FOXK1, LC3-II, P62, E-cadherin and MMP9 levels in lung tissue (scale bar, 100 μm and 50 μm). ( F ) MGC803 cells transfected with LV-shFOXK1-1 and LV-shFOXK1-2 were orthotopically transplanted into the stomach walls of nude mice (n=5). ( G , H ) Weight of gastric wall tumors (g) and number of metastatic tumors ( H ) at 28 days post transplantation with MGC803 cells. ( I , J ) Representative images ( I ) and number ( J ) of diffuse liver metastases. * P

    Journal: Aging (Albany NY)

    Article Title: Inhibiting Forkhead box K1 induces autophagy to reverse epithelial-mesenchymal transition and metastasis in gastric cancer by regulating Myc-associated zinc finger protein in an acidic microenvironment

    doi: 10.18632/aging.103013

    Figure Lengend Snippet: Downregulation of FOXK1 inhibits metastasis in vivo. ( A ) Lung tissues were obtained from nude mice injected with differently treated MGC803 cells (n=5). ( B ) The number of lung nodules per nude mouse was determined. ( C ) The lung weight of each nude mouse was calculated. ( D , E ) Representative images of H E staining of lung sections (scale bar, 100 μm and 50 μm) and IHC staining of FOXK1, LC3-II, P62, E-cadherin and MMP9 levels in lung tissue (scale bar, 100 μm and 50 μm). ( F ) MGC803 cells transfected with LV-shFOXK1-1 and LV-shFOXK1-2 were orthotopically transplanted into the stomach walls of nude mice (n=5). ( G , H ) Weight of gastric wall tumors (g) and number of metastatic tumors ( H ) at 28 days post transplantation with MGC803 cells. ( I , J ) Representative images ( I ) and number ( J ) of diffuse liver metastases. * P

    Article Snippet: Antibodies against FOXK1 (ab18196) and MMP-9 (ab38898) were purchased from Abcam (Cambridge, MA, USA), and antibodies against MAZ (NB100-86984) were purchased from Novus (Columbus, MO, USA).

    Techniques: In Vivo, Mouse Assay, Injection, Staining, Immunohistochemistry, Transfection, Transplantation Assay

    The silencing of FOXK1 induces autophagy to inhibit the migration and invasion of acidic GC cells. ( A – H ) Acidic MGC803 and AGS cells were infected with an empty lentivirus control (LV-ctrl) or with LV shFOXK1-1 or shFOXK1-2 and subsequently incubated for 24 h prior to pretreatment with 2 mM 3-MA or PBS (control). ( A , B ) Laser confocal microscopy analysis and quantification of transfected MGC803 cells with plasmid constructs containing LC3, which was fused with tandem mRFP-GFP tags. Scale bar, 10 μm. ( C , D ) The number of autophagosomes was observed and quantified under a transmission electron microscope. Scale bar, 2 μm. ( E ) Western blotting analysis of the levels of FOXK1, LC3-I, LC3-II, P62, Beclin1 and MMP9; β-actin was used as a loading control. ( F ) Matrigel cell invasion assays of MGC803 and AGS cells were performed, and the invading cells were quantified. ( G , H ) Scratch test evaluation of MGC803 cells. The wound healing area was analyzed using ImageJ software. Scale bar, 500 μm. The data are presented as the means ± S.D.s from three independent experiments. * P

    Journal: Aging (Albany NY)

    Article Title: Inhibiting Forkhead box K1 induces autophagy to reverse epithelial-mesenchymal transition and metastasis in gastric cancer by regulating Myc-associated zinc finger protein in an acidic microenvironment

    doi: 10.18632/aging.103013

    Figure Lengend Snippet: The silencing of FOXK1 induces autophagy to inhibit the migration and invasion of acidic GC cells. ( A – H ) Acidic MGC803 and AGS cells were infected with an empty lentivirus control (LV-ctrl) or with LV shFOXK1-1 or shFOXK1-2 and subsequently incubated for 24 h prior to pretreatment with 2 mM 3-MA or PBS (control). ( A , B ) Laser confocal microscopy analysis and quantification of transfected MGC803 cells with plasmid constructs containing LC3, which was fused with tandem mRFP-GFP tags. Scale bar, 10 μm. ( C , D ) The number of autophagosomes was observed and quantified under a transmission electron microscope. Scale bar, 2 μm. ( E ) Western blotting analysis of the levels of FOXK1, LC3-I, LC3-II, P62, Beclin1 and MMP9; β-actin was used as a loading control. ( F ) Matrigel cell invasion assays of MGC803 and AGS cells were performed, and the invading cells were quantified. ( G , H ) Scratch test evaluation of MGC803 cells. The wound healing area was analyzed using ImageJ software. Scale bar, 500 μm. The data are presented as the means ± S.D.s from three independent experiments. * P

    Article Snippet: Antibodies against FOXK1 (ab18196) and MMP-9 (ab38898) were purchased from Abcam (Cambridge, MA, USA), and antibodies against MAZ (NB100-86984) were purchased from Novus (Columbus, MO, USA).

    Techniques: Migration, Infection, Incubation, Confocal Microscopy, Transfection, Plasmid Preparation, Construct, Transmission Assay, Microscopy, Western Blot, Software

    Dual inhibition of mTOR and FOXK1 enhances autophagy and leads to the synergistic transfer of acidic GC cells. ( A – G ) Acidic MGC803 cells were treated with DMSO or rapamycin (100 nM) or infected with ctrl, shFOXK1-1, shFOXK1-2 or a combination of these for 24 h. ( A ) The red-only and yellow puncta of MGC803 cells transfected with mRFP-GFP-LC3 were observed under laser confocal conditions, and the quantitative results are shown in ( B ). Scale bar, 10 μm. ( C ) Western blotting was performed to assess the expression intensity of LC3-I, LC3-II, P62, E-cadherin, N-cadherin, Vimentin and MMP9. ( D – F ) The migration and invasive capabilities of MGC803 cells were examined. The invading cells are quantified in ( D , E ). The data are presented as the means ± S.D.s from three independent experiments. * P

    Journal: Aging (Albany NY)

    Article Title: Inhibiting Forkhead box K1 induces autophagy to reverse epithelial-mesenchymal transition and metastasis in gastric cancer by regulating Myc-associated zinc finger protein in an acidic microenvironment

    doi: 10.18632/aging.103013

    Figure Lengend Snippet: Dual inhibition of mTOR and FOXK1 enhances autophagy and leads to the synergistic transfer of acidic GC cells. ( A – G ) Acidic MGC803 cells were treated with DMSO or rapamycin (100 nM) or infected with ctrl, shFOXK1-1, shFOXK1-2 or a combination of these for 24 h. ( A ) The red-only and yellow puncta of MGC803 cells transfected with mRFP-GFP-LC3 were observed under laser confocal conditions, and the quantitative results are shown in ( B ). Scale bar, 10 μm. ( C ) Western blotting was performed to assess the expression intensity of LC3-I, LC3-II, P62, E-cadherin, N-cadherin, Vimentin and MMP9. ( D – F ) The migration and invasive capabilities of MGC803 cells were examined. The invading cells are quantified in ( D , E ). The data are presented as the means ± S.D.s from three independent experiments. * P

    Article Snippet: Antibodies against FOXK1 (ab18196) and MMP-9 (ab38898) were purchased from Abcam (Cambridge, MA, USA), and antibodies against MAZ (NB100-86984) were purchased from Novus (Columbus, MO, USA).

    Techniques: Inhibition, Infection, Transfection, Western Blot, Expressing, Migration

    Model of melatonin regulation of breast cancer cell invasion . Ligand-dependent activation of the membrane-associated G-protein-coupled receptor MT1 leads to coupling of G i2 protein to MT1 receptor. As a result, the Gα i2 subunit dissociates from the Gβγ subunits and inhibits the activity of adenylcyclase (AC), and this leads to a decrease in the intracellular level of cAMP and inhibition of protein kinase A (PKA) activity. The cAMP/PKA pathway cross-talks with the p38 pathway through PKA. In response to the reduced cAMP level, activity of p38 is suppressed, and this causes further downregulation of MMP-9 expression via repression of ETS1 transcriptional activity and, potentially, downregulation of MMP-2 transcription. MMP, matrix metalloproteinase.

    Journal: Breast Cancer Research : BCR

    Article Title: Inhibition of breast cancer cell invasion by melatonin is mediated through regulation of the p38 mitogen-activated protein kinase signaling pathway

    doi: 10.1186/bcr2794

    Figure Lengend Snippet: Model of melatonin regulation of breast cancer cell invasion . Ligand-dependent activation of the membrane-associated G-protein-coupled receptor MT1 leads to coupling of G i2 protein to MT1 receptor. As a result, the Gα i2 subunit dissociates from the Gβγ subunits and inhibits the activity of adenylcyclase (AC), and this leads to a decrease in the intracellular level of cAMP and inhibition of protein kinase A (PKA) activity. The cAMP/PKA pathway cross-talks with the p38 pathway through PKA. In response to the reduced cAMP level, activity of p38 is suppressed, and this causes further downregulation of MMP-9 expression via repression of ETS1 transcriptional activity and, potentially, downregulation of MMP-2 transcription. MMP, matrix metalloproteinase.

    Article Snippet: For MMP-2 and MMP-9 expression studies, the enriched conditioned medium from each treatment group was electrophoretically separated on a 10% SDS-polyacrylamide gel, and the blots were probed with anti-MMP-2 and anti-MMP-9 antibodies (Chemicon, now part of Millipore Corporation).

    Techniques: Activation Assay, Activity Assay, Inhibition, Expressing

    Melatonin suppresses the expression and activity of MMP-2 and MMP-9 in human breast cancer cells . Conditioned medium was collected and concentrated as described in Materials and methods. (a) Melatonin's effect on the protein expression of MMP-2 and MMP-9. MCF-7/6 cells were treated with melatonin (Mlt, 10 -9 M) or diluent (Control, 0.00001% ethanol) for 48 hours. Expression of MMP-2 and MMP-9 in the conditioned medium was analyzed by Western blot analysis using anti-MMP-2 and anti-MMP-9 antibody. (b) The effect of melatonin on the activity of MMP-2 and MMP-9. MCF-7/6 cells were treated with melatonin (Mlt, 10 -9 M) or diluent (Control, 0.00001% ethanol) for 48 hours. The gelatinase activity of MMP-9 and MMP-2 in the conditioned medium was determined by gelatin zymography. The band intensities of MMP-9 and MMP-2, respectively, are presented in the graph as percentages of control (set as 100%). * P

    Journal: Breast Cancer Research : BCR

    Article Title: Inhibition of breast cancer cell invasion by melatonin is mediated through regulation of the p38 mitogen-activated protein kinase signaling pathway

    doi: 10.1186/bcr2794

    Figure Lengend Snippet: Melatonin suppresses the expression and activity of MMP-2 and MMP-9 in human breast cancer cells . Conditioned medium was collected and concentrated as described in Materials and methods. (a) Melatonin's effect on the protein expression of MMP-2 and MMP-9. MCF-7/6 cells were treated with melatonin (Mlt, 10 -9 M) or diluent (Control, 0.00001% ethanol) for 48 hours. Expression of MMP-2 and MMP-9 in the conditioned medium was analyzed by Western blot analysis using anti-MMP-2 and anti-MMP-9 antibody. (b) The effect of melatonin on the activity of MMP-2 and MMP-9. MCF-7/6 cells were treated with melatonin (Mlt, 10 -9 M) or diluent (Control, 0.00001% ethanol) for 48 hours. The gelatinase activity of MMP-9 and MMP-2 in the conditioned medium was determined by gelatin zymography. The band intensities of MMP-9 and MMP-2, respectively, are presented in the graph as percentages of control (set as 100%). * P

    Article Snippet: For MMP-2 and MMP-9 expression studies, the enriched conditioned medium from each treatment group was electrophoretically separated on a 10% SDS-polyacrylamide gel, and the blots were probed with anti-MMP-2 and anti-MMP-9 antibodies (Chemicon, now part of Millipore Corporation).

    Techniques: Expressing, Activity Assay, Western Blot, Zymography

    Biochemical characterization of the osteoclast-likeness of the various MGCs. (A) Cell lysates were taken at indicated time points during the differentiation protocol and analyzed for MMP-9 by Western blotting. 1* indicates lysates from RAW 264.7 cells prior to LPS/IFNγ or IL-4 addition. Numbers below MMP-9 blot represent densitometry readings for MMP-9 expression with 1.0 being day 2, RANKL-treated cells. (B) Cell lysates were taken from day 2 – day 4 during the differentiation protocol and analyzed for Cathepsin K expression. The last lane is the cell lysate from day 4 untreated RAW 264.7 cells. (C) Same as (B) but blots were analyzed for Arginase and iNOS (arrow). GAPDH was used as a loading control. Blots are representative of three independent experiments.

    Journal: PLoS ONE

    Article Title: Modulation of Osteoclastogenesis with Macrophage M1- and M2-Inducing Stimuli

    doi: 10.1371/journal.pone.0104498

    Figure Lengend Snippet: Biochemical characterization of the osteoclast-likeness of the various MGCs. (A) Cell lysates were taken at indicated time points during the differentiation protocol and analyzed for MMP-9 by Western blotting. 1* indicates lysates from RAW 264.7 cells prior to LPS/IFNγ or IL-4 addition. Numbers below MMP-9 blot represent densitometry readings for MMP-9 expression with 1.0 being day 2, RANKL-treated cells. (B) Cell lysates were taken from day 2 – day 4 during the differentiation protocol and analyzed for Cathepsin K expression. The last lane is the cell lysate from day 4 untreated RAW 264.7 cells. (C) Same as (B) but blots were analyzed for Arginase and iNOS (arrow). GAPDH was used as a loading control. Blots are representative of three independent experiments.

    Article Snippet: Rabbit anti-MMP-9 and rabbit anti-cathepsin K antibodies were purchased from Abcam (Toronto, ON).

    Techniques: Western Blot, Expressing

    MMP-9 mediates Slug-dependent EMT in NRK52 cells downstream of TGF-β1. A–F: Immunofluorescence images showing E-cadherin ( A and B: Scale bar = 15 μm), α-SMA ( C and D) and β-catenin ( E and F ) of NRK52e cells

    Journal: The American Journal of Pathology

    Article Title: Disruption of E-Cadherin by Matrix Metalloproteinase Directly Mediates Epithelial-Mesenchymal Transition Downstream of Transforming Growth Factor-?1 in Renal Tubular Epithelial Cells

    doi: 10.2353/ajpath.2009.080983

    Figure Lengend Snippet: MMP-9 mediates Slug-dependent EMT in NRK52 cells downstream of TGF-β1. A–F: Immunofluorescence images showing E-cadherin ( A and B: Scale bar = 15 μm), α-SMA ( C and D) and β-catenin ( E and F ) of NRK52e cells

    Article Snippet: Anti-E-cadherin antibody (BD Transduction Lab, Lexington, KY), anti-α-smooth muscle actin (SMA) (Chemicon, Billerica, MA), anti-β-catenin antibody (BD Transduction Lab), anti-Slug antibody (Santa Cruz Biotechnology, St Louis, MO), anti-β-actin antibody (Sigma), anti-MMP-9 antibody (Calbiochem) and anti-HSP47 antibody (Santa Cruz Biotechnology) were used with respective secondary antibodies: fluorescein isothiocyanate-conjugated rat anti-mouse IgG2a/b (BD Biosciences PharMingen, San Jose, CA), Texas Red conjugated goat anti-rabbit IgGH/L (Calbiochem),, or biotin conjugated rabbit anti-mouse IgG1 (Zymed, San Francisco, CA) together with fluorescein-conjugated streptavidin (eBioscience, San Diego, CA).

    Techniques: Immunofluorescence

    MMPs up-regulation and EMT in diseased kidney. A: Up-regulation of MMP-2, MMP-3 and MMP-9 in kidney of rats with AN at 4 weeks by immunohistochemistry. Arrows indicate the tubular and interstitial staining (dark brown) of corresponding MMPs. B: Immunohistochemical

    Journal: The American Journal of Pathology

    Article Title: Disruption of E-Cadherin by Matrix Metalloproteinase Directly Mediates Epithelial-Mesenchymal Transition Downstream of Transforming Growth Factor-?1 in Renal Tubular Epithelial Cells

    doi: 10.2353/ajpath.2009.080983

    Figure Lengend Snippet: MMPs up-regulation and EMT in diseased kidney. A: Up-regulation of MMP-2, MMP-3 and MMP-9 in kidney of rats with AN at 4 weeks by immunohistochemistry. Arrows indicate the tubular and interstitial staining (dark brown) of corresponding MMPs. B: Immunohistochemical

    Article Snippet: Anti-E-cadherin antibody (BD Transduction Lab, Lexington, KY), anti-α-smooth muscle actin (SMA) (Chemicon, Billerica, MA), anti-β-catenin antibody (BD Transduction Lab), anti-Slug antibody (Santa Cruz Biotechnology, St Louis, MO), anti-β-actin antibody (Sigma), anti-MMP-9 antibody (Calbiochem) and anti-HSP47 antibody (Santa Cruz Biotechnology) were used with respective secondary antibodies: fluorescein isothiocyanate-conjugated rat anti-mouse IgG2a/b (BD Biosciences PharMingen, San Jose, CA), Texas Red conjugated goat anti-rabbit IgGH/L (Calbiochem),, or biotin conjugated rabbit anti-mouse IgG1 (Zymed, San Francisco, CA) together with fluorescein-conjugated streptavidin (eBioscience, San Diego, CA).

    Techniques: Immunohistochemistry, Staining

    Co-treatment of Zyflamend and cisplatin decreased cell proliferation, increased apoptosis, and suppressed NFκB signaling in T24R xenografts. Notes: ( A ) Representative IHC staining of proliferative marker (Ki-67), apoptosis marker (cleaved caspase 3), RelA, and MMP9 from the sections of vehicle, cisplatin, and/or Zyflamend treatment groups. Quantification of Ki-67-positive cancer cells ( B ) and cleaved caspase 3 ( C ) in the sections of T24R xenografts with different treatments. Data are presented as mean ± SE; * P

    Journal: OncoTargets and therapy

    Article Title: Combination chemotherapy with Zyflamend reduced the acquired resistance of bladder cancer cells to cisplatin through inhibiting NFκB signaling pathway

    doi: 10.2147/OTT.S162255

    Figure Lengend Snippet: Co-treatment of Zyflamend and cisplatin decreased cell proliferation, increased apoptosis, and suppressed NFκB signaling in T24R xenografts. Notes: ( A ) Representative IHC staining of proliferative marker (Ki-67), apoptosis marker (cleaved caspase 3), RelA, and MMP9 from the sections of vehicle, cisplatin, and/or Zyflamend treatment groups. Quantification of Ki-67-positive cancer cells ( B ) and cleaved caspase 3 ( C ) in the sections of T24R xenografts with different treatments. Data are presented as mean ± SE; * P

    Article Snippet: For IHC staining, sections were stained by the antibodies against Ki-67 (M7240, 1:1,000; DAKO), cleaved caspase 3 (9661, 1:1,000; Cell Signaling Technologies), RelA (8242, 1:1,000; Cell Signaling Technologies), and MMP9 (13667, 1:1,000; Cell Signaling Technologies) following the standard protocol.

    Techniques: Immunohistochemistry, Staining, Marker

    Elimination of SnCs by ABT263 and alleviation inflammatory microenvironment in vivo. ( A ) immunohistochemical analysis for expression of p16 INK4a , HMGB1, COLII and ACAN of rat cartilage after 2 weeks intra-articular injection of ABT263. Result from ABT263 of 1.0 mM was presented. Scale bar: 100 μm. ( B ) mRNA level analysis using real-time qPCR for CDKN1A, CDKN2A, MMP13, ADAMTS5, IL1β, IL6, COLII and ACAN in the rat knee joint. Data are shown as mean ± standard deviation. N = 3 per group. +p

    Journal: Aging (Albany NY)

    Article Title: Navitoclax (ABT263) reduces inflammation and promotes chondrogenic phenotype by clearing senescent osteoarthritic chondrocytes in osteoarthritis

    doi: 10.18632/aging.103177

    Figure Lengend Snippet: Elimination of SnCs by ABT263 and alleviation inflammatory microenvironment in vivo. ( A ) immunohistochemical analysis for expression of p16 INK4a , HMGB1, COLII and ACAN of rat cartilage after 2 weeks intra-articular injection of ABT263. Result from ABT263 of 1.0 mM was presented. Scale bar: 100 μm. ( B ) mRNA level analysis using real-time qPCR for CDKN1A, CDKN2A, MMP13, ADAMTS5, IL1β, IL6, COLII and ACAN in the rat knee joint. Data are shown as mean ± standard deviation. N = 3 per group. +p

    Article Snippet: Sections were incubated overnight at 4 °C with the following primary antibodies: p16INK4a (1:500; Abcam, ab54210), MMP-13 (1:200; Abcam, ab39012), type II collagen (1:300; Abcam, ab34712) and HMGB1 (1:500; Abcam, ab18256).

    Techniques: In Vivo, Immunohistochemistry, Expressing, Injection, Real-time Polymerase Chain Reaction, Standard Deviation

    Effect of ABT263 on OA chondrocytes in monolayer culture. ( A ) qPCR analysis of mRNA expression for MMP13, ADAMTS5, IL1β, IL6, COLII, ACAN and SOX9 at Day 3 and Day 10 separately. ( B ) Immunoblotting analysis of protein level for MMP13, ADAMTS5, IL1β, COLII, ACAN and SOX9 at Day 3 and Day 10 separately. Data are shown as mean ± standard deviation. N = 3 per group.

    Journal: Aging (Albany NY)

    Article Title: Navitoclax (ABT263) reduces inflammation and promotes chondrogenic phenotype by clearing senescent osteoarthritic chondrocytes in osteoarthritis

    doi: 10.18632/aging.103177

    Figure Lengend Snippet: Effect of ABT263 on OA chondrocytes in monolayer culture. ( A ) qPCR analysis of mRNA expression for MMP13, ADAMTS5, IL1β, IL6, COLII, ACAN and SOX9 at Day 3 and Day 10 separately. ( B ) Immunoblotting analysis of protein level for MMP13, ADAMTS5, IL1β, COLII, ACAN and SOX9 at Day 3 and Day 10 separately. Data are shown as mean ± standard deviation. N = 3 per group.

    Article Snippet: Sections were incubated overnight at 4 °C with the following primary antibodies: p16INK4a (1:500; Abcam, ab54210), MMP-13 (1:200; Abcam, ab39012), type II collagen (1:300; Abcam, ab34712) and HMGB1 (1:500; Abcam, ab18256).

    Techniques: Real-time Polymerase Chain Reaction, Expressing, Standard Deviation

    Improvement of inflammation and promotion of proteoglycan anabolism. ( A ) immunohistochemical analysis of senescence and inflammation-related markers and safranine O staining after 21-day pellet culture. Scale bar: 200 μm. ( B ) mRNA expression analysis by qPCR for CDKN1A, CDKN2A, MMP13, ADAMTS5, IL1β, IL6, COLII, ACAN and SOX9. ( C ) protein level analysis using Immunoblotting for C-Caspase3, p21, MMP13, ADAMTS5, IL1β, COLII, ACAN and SOX9. Data are shown as mean ± standard deviation. N = 3 per group.

    Journal: Aging (Albany NY)

    Article Title: Navitoclax (ABT263) reduces inflammation and promotes chondrogenic phenotype by clearing senescent osteoarthritic chondrocytes in osteoarthritis

    doi: 10.18632/aging.103177

    Figure Lengend Snippet: Improvement of inflammation and promotion of proteoglycan anabolism. ( A ) immunohistochemical analysis of senescence and inflammation-related markers and safranine O staining after 21-day pellet culture. Scale bar: 200 μm. ( B ) mRNA expression analysis by qPCR for CDKN1A, CDKN2A, MMP13, ADAMTS5, IL1β, IL6, COLII, ACAN and SOX9. ( C ) protein level analysis using Immunoblotting for C-Caspase3, p21, MMP13, ADAMTS5, IL1β, COLII, ACAN and SOX9. Data are shown as mean ± standard deviation. N = 3 per group.

    Article Snippet: Sections were incubated overnight at 4 °C with the following primary antibodies: p16INK4a (1:500; Abcam, ab54210), MMP-13 (1:200; Abcam, ab39012), type II collagen (1:300; Abcam, ab34712) and HMGB1 (1:500; Abcam, ab18256).

    Techniques: Immunohistochemistry, Staining, Expressing, Real-time Polymerase Chain Reaction, Standard Deviation

    Notch1 effects MT1-MMP-dependent melanoma cell growth. A , expression levels of MT1-MMP ( MT1 ) and Notch1 NIC in WM266-4 cells expressing shGFP or shMT1-MMP and transduced with active Notch1 ( NIC ). β- act , β-actin. B , relative growth at a

    Journal: The Journal of Biological Chemistry

    Article Title: Noncanonical Activation of Notch1 Protein by Membrane Type 1 Matrix Metalloproteinase (MT1-MMP) Controls Melanoma Cell Proliferation *

    doi: 10.1074/jbc.M113.516039

    Figure Lengend Snippet: Notch1 effects MT1-MMP-dependent melanoma cell growth. A , expression levels of MT1-MMP ( MT1 ) and Notch1 NIC in WM266-4 cells expressing shGFP or shMT1-MMP and transduced with active Notch1 ( NIC ). β- act , β-actin. B , relative growth at a

    Article Snippet: Membranes were probed with the following antibodies: anti-Notch1-TM (C20, Santa Cruz Biotechnology, Santa Cruz, CA); anti-Notch1NIC (Val-1744) (Cell Signaling Technologies, Beverly, MA); anti MT1-MMP (clone LEM-2/15.8, Millipore, Billerica, MA); anti-ADAM10 (Abcam, Cambridge, MA); and anti-TACE (tumor necrosis factor-α-converting enzyme) (ADAM17) (eBioscience, San Diego, CA).

    Techniques: Expressing, Transduction, Activated Clotting Time Assay

    Notch1 NIC and MT1-MMP expression correlate in melanoma. A , Western blotting on nine flash-frozen, human melanoma tumors showing expression of ADAM17 ( A-17 ), ADAM10 ( A-10 ), MT1-MMP ( MT1 ), and active Notch1 ( NIC ). β-Actin (β- Act ) was used

    Journal: The Journal of Biological Chemistry

    Article Title: Noncanonical Activation of Notch1 Protein by Membrane Type 1 Matrix Metalloproteinase (MT1-MMP) Controls Melanoma Cell Proliferation *

    doi: 10.1074/jbc.M113.516039

    Figure Lengend Snippet: Notch1 NIC and MT1-MMP expression correlate in melanoma. A , Western blotting on nine flash-frozen, human melanoma tumors showing expression of ADAM17 ( A-17 ), ADAM10 ( A-10 ), MT1-MMP ( MT1 ), and active Notch1 ( NIC ). β-Actin (β- Act ) was used

    Article Snippet: Membranes were probed with the following antibodies: anti-Notch1-TM (C20, Santa Cruz Biotechnology, Santa Cruz, CA); anti-Notch1NIC (Val-1744) (Cell Signaling Technologies, Beverly, MA); anti MT1-MMP (clone LEM-2/15.8, Millipore, Billerica, MA); anti-ADAM10 (Abcam, Cambridge, MA); and anti-TACE (tumor necrosis factor-α-converting enzyme) (ADAM17) (eBioscience, San Diego, CA).

    Techniques: Expressing, Western Blot, Activated Clotting Time Assay

    Active MT1-MMP promotes Notch1 cleavage independently of ADAM10 or -17. A , MT1-MMP ( MT1 ) protein expression in the syngeneic cell lines WM115 (primary melanoma) and WM2660-4 (metastatic melanoma). β- act , β-actin. B , schematic representation

    Journal: The Journal of Biological Chemistry

    Article Title: Noncanonical Activation of Notch1 Protein by Membrane Type 1 Matrix Metalloproteinase (MT1-MMP) Controls Melanoma Cell Proliferation *

    doi: 10.1074/jbc.M113.516039

    Figure Lengend Snippet: Active MT1-MMP promotes Notch1 cleavage independently of ADAM10 or -17. A , MT1-MMP ( MT1 ) protein expression in the syngeneic cell lines WM115 (primary melanoma) and WM2660-4 (metastatic melanoma). β- act , β-actin. B , schematic representation

    Article Snippet: Membranes were probed with the following antibodies: anti-Notch1-TM (C20, Santa Cruz Biotechnology, Santa Cruz, CA); anti-Notch1NIC (Val-1744) (Cell Signaling Technologies, Beverly, MA); anti MT1-MMP (clone LEM-2/15.8, Millipore, Billerica, MA); anti-ADAM10 (Abcam, Cambridge, MA); and anti-TACE (tumor necrosis factor-α-converting enzyme) (ADAM17) (eBioscience, San Diego, CA).

    Techniques: Expressing, Activated Clotting Time Assay

    MT1-MMP affects melanoma cell growth. A , relative growth of WM266-4 expressing a control shRNA ( shGFP ) or shMT1-MMP. B , relative growth of WM115 expressing a control vector ( pLM ) or either active ( MT1-FL ) or inactive ( MT1-Mut ) MT1-MMP. *, p

    Journal: The Journal of Biological Chemistry

    Article Title: Noncanonical Activation of Notch1 Protein by Membrane Type 1 Matrix Metalloproteinase (MT1-MMP) Controls Melanoma Cell Proliferation *

    doi: 10.1074/jbc.M113.516039

    Figure Lengend Snippet: MT1-MMP affects melanoma cell growth. A , relative growth of WM266-4 expressing a control shRNA ( shGFP ) or shMT1-MMP. B , relative growth of WM115 expressing a control vector ( pLM ) or either active ( MT1-FL ) or inactive ( MT1-Mut ) MT1-MMP. *, p

    Article Snippet: Membranes were probed with the following antibodies: anti-Notch1-TM (C20, Santa Cruz Biotechnology, Santa Cruz, CA); anti-Notch1NIC (Val-1744) (Cell Signaling Technologies, Beverly, MA); anti MT1-MMP (clone LEM-2/15.8, Millipore, Billerica, MA); anti-ADAM10 (Abcam, Cambridge, MA); and anti-TACE (tumor necrosis factor-α-converting enzyme) (ADAM17) (eBioscience, San Diego, CA).

    Techniques: Expressing, shRNA, Plasmid Preparation

    MT1-MMP inhibition reduces Notch1 cleavage. A , expression of full-length Notch1 ( N1 ), Notch1 NIC ( NIC ), MT1-MMP ( MT1 ), and MMP2 in WM266-4 cells expressing shRNAs against GFP, MT1-MMP, MMP2, or both MT1-MMP and MMP2. The ratio between the Notch1 NIC band

    Journal: The Journal of Biological Chemistry

    Article Title: Noncanonical Activation of Notch1 Protein by Membrane Type 1 Matrix Metalloproteinase (MT1-MMP) Controls Melanoma Cell Proliferation *

    doi: 10.1074/jbc.M113.516039

    Figure Lengend Snippet: MT1-MMP inhibition reduces Notch1 cleavage. A , expression of full-length Notch1 ( N1 ), Notch1 NIC ( NIC ), MT1-MMP ( MT1 ), and MMP2 in WM266-4 cells expressing shRNAs against GFP, MT1-MMP, MMP2, or both MT1-MMP and MMP2. The ratio between the Notch1 NIC band

    Article Snippet: Membranes were probed with the following antibodies: anti-Notch1-TM (C20, Santa Cruz Biotechnology, Santa Cruz, CA); anti-Notch1NIC (Val-1744) (Cell Signaling Technologies, Beverly, MA); anti MT1-MMP (clone LEM-2/15.8, Millipore, Billerica, MA); anti-ADAM10 (Abcam, Cambridge, MA); and anti-TACE (tumor necrosis factor-α-converting enzyme) (ADAM17) (eBioscience, San Diego, CA).

    Techniques: Inhibition, Expressing

    Upregulation of MMP-9 expression by CS. Notes: Immunohistochemical detection of MMP-9 (DAB, brown) in ( A ) CS-exposed mice and ( B ) air-exposed mice. Sections were counterstained with Mayer hematoxylin (blue). Scale bar=50 µm. ( C ) Number of MMP-9–positive cells per square millimeter, n=5. ( D ) MMP-9 mRNA level in the lung tissues, n=4. ( E ) Co-immunofluorescent staining for MMP-9 (AlexaFluor 594, red) and F4/80 (AlexaFluor 488, green) in the lungs of CS-exposed mice. Nuclei were stained with DAPI (blue). ( F and G ) Representative Western blot and relative quantification normalized to β-actin for MMP-9 in MH-S stimulated with CSE for 48 hours; n=3. Arrows indicate positive cells. Bars indicate the mean value, and error bars indicate SEM. * P

    Journal: International Journal of Chronic Obstructive Pulmonary Disease

    Article Title: Cigarette smoke-induced RANKL expression enhances MMP-9 production by alveolar macrophages

    doi: 10.2147/COPD.S190023

    Figure Lengend Snippet: Upregulation of MMP-9 expression by CS. Notes: Immunohistochemical detection of MMP-9 (DAB, brown) in ( A ) CS-exposed mice and ( B ) air-exposed mice. Sections were counterstained with Mayer hematoxylin (blue). Scale bar=50 µm. ( C ) Number of MMP-9–positive cells per square millimeter, n=5. ( D ) MMP-9 mRNA level in the lung tissues, n=4. ( E ) Co-immunofluorescent staining for MMP-9 (AlexaFluor 594, red) and F4/80 (AlexaFluor 488, green) in the lungs of CS-exposed mice. Nuclei were stained with DAPI (blue). ( F and G ) Representative Western blot and relative quantification normalized to β-actin for MMP-9 in MH-S stimulated with CSE for 48 hours; n=3. Arrows indicate positive cells. Bars indicate the mean value, and error bars indicate SEM. * P

    Article Snippet: Afterward, the sections were treated with goat serum (ZLI-6056; ZSGB-Bio, Beijing, China) and incubated overnight with MMP-9 antibody (Abcam, Cambridge, UK).

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

    MMP-9 expression was regulated by RANKL. Notes: ( A ) MMP-9 mRNA was upregulated in MH-S under RANKL stimulation. ( B and C ) MMP-12 and TIMP1 mRNA in MH-S under RANKL stimulation. The ratio of ( D ) MMP-9 mRNA to TIMP1 mRNA and ( E ) MMP-12 mRNA to TIMP1 mRNA under RANKL stimulation. ( F and G ) Representative Western blot and relative quantification normalized to β-actin for MMP-9 in MH-S stimulated with RANKL. ( H and I ) Representative Western blot and relative quantification normalized to β-actin for MMP-9 in MH-S stimulated with combined CSE with monoclonal anti-RANKL antibody. n=3. Bars indicate the mean value, and error bars indicate SEM. * P

    Journal: International Journal of Chronic Obstructive Pulmonary Disease

    Article Title: Cigarette smoke-induced RANKL expression enhances MMP-9 production by alveolar macrophages

    doi: 10.2147/COPD.S190023

    Figure Lengend Snippet: MMP-9 expression was regulated by RANKL. Notes: ( A ) MMP-9 mRNA was upregulated in MH-S under RANKL stimulation. ( B and C ) MMP-12 and TIMP1 mRNA in MH-S under RANKL stimulation. The ratio of ( D ) MMP-9 mRNA to TIMP1 mRNA and ( E ) MMP-12 mRNA to TIMP1 mRNA under RANKL stimulation. ( F and G ) Representative Western blot and relative quantification normalized to β-actin for MMP-9 in MH-S stimulated with RANKL. ( H and I ) Representative Western blot and relative quantification normalized to β-actin for MMP-9 in MH-S stimulated with combined CSE with monoclonal anti-RANKL antibody. n=3. Bars indicate the mean value, and error bars indicate SEM. * P

    Article Snippet: Afterward, the sections were treated with goat serum (ZLI-6056; ZSGB-Bio, Beijing, China) and incubated overnight with MMP-9 antibody (Abcam, Cambridge, UK).

    Techniques: Expressing, Western Blot

    The schematic diagram of OA treatment by TGF-β3 and BMP7. ( A ) Osteoarthritic cartilage with a degenerative superficial layer and loss of chondrocytes shows higher catabolism, reflected by elevated MMP-1/3/13 expression; ( B ) After treatment with the combination of TGF-β3 and BMP7, the superficial layer with chondrocytes with newly gained energy may exhibit higher anabolism, reflected by increased expression of COL2A1 and ACAN .

    Journal: Scientific Reports

    Article Title: Co-treatment of TGF-β3 and BMP7 is superior in stimulating chondrocyte redifferentiation in both hypoxia and normoxia compared to single treatments

    doi: 10.1038/s41598-018-27602-y

    Figure Lengend Snippet: The schematic diagram of OA treatment by TGF-β3 and BMP7. ( A ) Osteoarthritic cartilage with a degenerative superficial layer and loss of chondrocytes shows higher catabolism, reflected by elevated MMP-1/3/13 expression; ( B ) After treatment with the combination of TGF-β3 and BMP7, the superficial layer with chondrocytes with newly gained energy may exhibit higher anabolism, reflected by increased expression of COL2A1 and ACAN .

    Article Snippet: The content of MMP1 dissolved in medium was measured by ELISA using a mouse anti-human MMP1 antibody (MAB901-SP, R & D systems) as described .

    Techniques: Expressing

    Adtrp deficiency leads to increased expression of matrix metallopeptidase‐9 (Mmp9) mRNA , protein, and activity levels in zebrafish embryos and newborn mice. A , Whole‐mount in situ hybridization for mmp9 (blue) in 72 hours postfertilization (hpf) control and adtrp zebrafish morphants. Blue arrow: specific signal accumulation. Black arrow: caudal vascular plexus ( CVP ). n, biological replicates. Bars, 100 μm. B , Immunofluorescence staining with anti‐Mmp‐9 (Cy3, red) IgG on cross‐sectioned CVP (green) of 72‐hpf Tg(fli1: EGFP )y1 ( EGFP , fluorescent vascular reporter) control and adtrp morphants±human ADTRP mRNA . Bars, 100 μm. C , Total lysates of 72‐hpf zebrafish embryos incubated ±100 μmol/L of pan‐ MMP inhibitor GM 6001 for 24 hours and analyzed by 10% gelatin gel electrophoresis. Left: representative zymogram (grayscale inverted image). Histogram: semiquantitative densitometry and statistics (mean± SEM by t test). * P

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

    Article Title: Role of ADTRP (Androgen‐Dependent Tissue Factor Pathway Inhibitor Regulating Protein) in Vascular Development and Function

    doi: 10.1161/JAHA.118.010690

    Figure Lengend Snippet: Adtrp deficiency leads to increased expression of matrix metallopeptidase‐9 (Mmp9) mRNA , protein, and activity levels in zebrafish embryos and newborn mice. A , Whole‐mount in situ hybridization for mmp9 (blue) in 72 hours postfertilization (hpf) control and adtrp zebrafish morphants. Blue arrow: specific signal accumulation. Black arrow: caudal vascular plexus ( CVP ). n, biological replicates. Bars, 100 μm. B , Immunofluorescence staining with anti‐Mmp‐9 (Cy3, red) IgG on cross‐sectioned CVP (green) of 72‐hpf Tg(fli1: EGFP )y1 ( EGFP , fluorescent vascular reporter) control and adtrp morphants±human ADTRP mRNA . Bars, 100 μm. C , Total lysates of 72‐hpf zebrafish embryos incubated ±100 μmol/L of pan‐ MMP inhibitor GM 6001 for 24 hours and analyzed by 10% gelatin gel electrophoresis. Left: representative zymogram (grayscale inverted image). Histogram: semiquantitative densitometry and statistics (mean± SEM by t test). * P

    Article Snippet: Antibodies Primary antibodies, used according to the manufacturers, were as follows: mouse monoclonal antibody (mAb) anti‐FLAG/DDK (TA50011; OriGene, Rockville, MD); rabbit mAb anti‐non‐phospho (active) β‐catenin (Ser33/37/Thr41; 8814; Cell Signaling Technology, Danvers, MA); rabbit anti‐β‐catenin (ab16051; Abcam, Cambridge, MA); rat mAb anti‐mouse CD31 (553370; BD Pharmingen, San Diego, CA); mouse mAb anti‐Fibrin II β‐chain (NYBT2G1; Accurate Chemical & Scientific, Westbury, NY); rabbit mAb anti‐desmin (1466‐1; Epitomics, Inc, Eugene, OR); rat mAb anti‐CD144 (cadherin‐5/VE‐cadherin [VEC]; 550548; BD Pharmingen); Alexa Fluor 488–labeled mouse mAb anti‐claudin‐5 (352588; Thermo Fisher Scientific, Waltham, MA); goat anti‐MMP‐9 (AF909; R & D Systems, Minneapolis, MN); mouse mAb anti‐MC tryptase (M7052; Dako, Carpinteria, CA); rabbit anti‐laminin (Z0097; Dako); rabbit anti‐mouse collagen‐IV (2150‐1470; Bio‐Rad, Hercules, CA); rabbit anti‐β‐galactosidase (β‐GAL; AB1211; EMD Millipore, Burlington, MA); rat mAb anti‐mouse erythroid cells (TER‐119; 116202; BioLegend, San Diego, CA); rabbit anti–platelet‐derived growth factor receptor beta (P20; sc‐339; Santa Cruz Biotechnology, Santa Cruz, CA); rabbit anti‐human ADTRP (HPA048113; Sigma‐Aldrich, St. Louis, MO); biotin‐labeled rat mAb anti‐mouse Ly6G/Ly‐6C (GR‐1; 108404; BioLegend); mouse mAb anti–enhanced green fluorescent protein (632381; Clontech Laboratories, Mountain View, CA); rabbit anti–glyceraldehyde 3‐phosphate dehydrogenase (G9545; Sigma‐Aldrich); and rabbit anti‐zebrafish Mmp‐9 (55345; AnaSpec, Fremont, CA).

    Techniques: Expressing, Activity Assay, Mouse Assay, In Situ Hybridization, Immunofluorescence, Staining, Incubation, Nucleic Acid Electrophoresis

    ADTRP  deficiency‐induced matrix metallopeptidase‐9 ( MMP ‐9) is regulated by canonical Wnt signaling.  A , Double immunofluorescence with anti‐ MMP ‐9 ( FITC , green) and anti‐β‐Galactosidase (β‐ GAL ; Cy3, red), and confocal microscopy in whole‐mount P 0  pups lung of  Adtrp −/− BAT ‐ GAL z+  and wild‐type ( WT )  BAT ‐ GAL z+ . Arrows: partial expression of β‐ GAL  (active β‐catenin reporter) in  MMP ‐9–producing cells. Blue, nuclei. Bars, 50 μm.  B ,  mRNA  expression of  Mmp9  in primary fibroblasts cultured from  WT  and  Adtrp −/−  P 0  pups cephalic skin explants. Gray bars: treatment with Wnt3a medium for 24 hours. Values are fold‐change from  WT  (arbitrarily set to 1.0) after correction for β ‐2 Microglobulin  as internal control. Data are mean± SEM  by 1‐way  ANOVA  with Bonferroni's multicomparison test. Markers represent mean values of 3 replicate cell cultures isolated from 2 mice each. ** P

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

    Article Title: Role of ADTRP (Androgen‐Dependent Tissue Factor Pathway Inhibitor Regulating Protein) in Vascular Development and Function

    doi: 10.1161/JAHA.118.010690

    Figure Lengend Snippet: ADTRP deficiency‐induced matrix metallopeptidase‐9 ( MMP ‐9) is regulated by canonical Wnt signaling. A , Double immunofluorescence with anti‐ MMP ‐9 ( FITC , green) and anti‐β‐Galactosidase (β‐ GAL ; Cy3, red), and confocal microscopy in whole‐mount P 0 pups lung of Adtrp −/− BAT ‐ GAL z+ and wild‐type ( WT ) BAT ‐ GAL z+ . Arrows: partial expression of β‐ GAL (active β‐catenin reporter) in MMP ‐9–producing cells. Blue, nuclei. Bars, 50 μm. B , mRNA expression of Mmp9 in primary fibroblasts cultured from WT and Adtrp −/− P 0 pups cephalic skin explants. Gray bars: treatment with Wnt3a medium for 24 hours. Values are fold‐change from WT (arbitrarily set to 1.0) after correction for β ‐2 Microglobulin as internal control. Data are mean± SEM by 1‐way ANOVA with Bonferroni's multicomparison test. Markers represent mean values of 3 replicate cell cultures isolated from 2 mice each. ** P

    Article Snippet: Antibodies Primary antibodies, used according to the manufacturers, were as follows: mouse monoclonal antibody (mAb) anti‐FLAG/DDK (TA50011; OriGene, Rockville, MD); rabbit mAb anti‐non‐phospho (active) β‐catenin (Ser33/37/Thr41; 8814; Cell Signaling Technology, Danvers, MA); rabbit anti‐β‐catenin (ab16051; Abcam, Cambridge, MA); rat mAb anti‐mouse CD31 (553370; BD Pharmingen, San Diego, CA); mouse mAb anti‐Fibrin II β‐chain (NYBT2G1; Accurate Chemical & Scientific, Westbury, NY); rabbit mAb anti‐desmin (1466‐1; Epitomics, Inc, Eugene, OR); rat mAb anti‐CD144 (cadherin‐5/VE‐cadherin [VEC]; 550548; BD Pharmingen); Alexa Fluor 488–labeled mouse mAb anti‐claudin‐5 (352588; Thermo Fisher Scientific, Waltham, MA); goat anti‐MMP‐9 (AF909; R & D Systems, Minneapolis, MN); mouse mAb anti‐MC tryptase (M7052; Dako, Carpinteria, CA); rabbit anti‐laminin (Z0097; Dako); rabbit anti‐mouse collagen‐IV (2150‐1470; Bio‐Rad, Hercules, CA); rabbit anti‐β‐galactosidase (β‐GAL; AB1211; EMD Millipore, Burlington, MA); rat mAb anti‐mouse erythroid cells (TER‐119; 116202; BioLegend, San Diego, CA); rabbit anti–platelet‐derived growth factor receptor beta (P20; sc‐339; Santa Cruz Biotechnology, Santa Cruz, CA); rabbit anti‐human ADTRP (HPA048113; Sigma‐Aldrich, St. Louis, MO); biotin‐labeled rat mAb anti‐mouse Ly6G/Ly‐6C (GR‐1; 108404; BioLegend); mouse mAb anti–enhanced green fluorescent protein (632381; Clontech Laboratories, Mountain View, CA); rabbit anti–glyceraldehyde 3‐phosphate dehydrogenase (G9545; Sigma‐Aldrich); and rabbit anti‐zebrafish Mmp‐9 (55345; AnaSpec, Fremont, CA).

    Techniques: Immunofluorescence, Confocal Microscopy, Expressing, Cell Culture, Isolation, Mouse Assay

    MMP-9 is involved in TGM2-mediated migration and invasion in the TRAIL-resistant cells. A , A549-TR cells were transfected with negative control siRNA or TGM2-siRNA and incubated for 48 h. Equal amounts of cell extracts were detected for TGM2, MMP-1, MMP-2,

    Journal: The Journal of Biological Chemistry

    Article Title: Epidermal Growth Factor Receptor-mediated Tissue Transglutaminase Overexpression Couples Acquired Tumor Necrosis Factor-related Apoptosis-inducing Ligand Resistance and Migration through c-FLIP and MMP-9 Proteins in Lung Cancer Cells *

    doi: 10.1074/jbc.M110.207571

    Figure Lengend Snippet: MMP-9 is involved in TGM2-mediated migration and invasion in the TRAIL-resistant cells. A , A549-TR cells were transfected with negative control siRNA or TGM2-siRNA and incubated for 48 h. Equal amounts of cell extracts were detected for TGM2, MMP-1, MMP-2,

    Article Snippet: The following antibodies were used for Western blot: anti-TGM2 (Santa Cruz Biotechnology), anti-β-tubulin and β-actin (Sigma-Aldrich); anti-MMP-1, MMP-2, and MMP-9 (Calbiochem); anti-c-FLIP and Mcl-1 (Alexis Laboratories); anti-phospho-EGFR (Y1086, Abcam), anti-EGFR, anti-ERK, anti-Akt, and anti-phospho-Akt (Ser-473) (Cell Signaling Technology); anti-phospho-ERK and phospho-JNK (BIOSOURCE); and anti-JNK1 (BD Biosciences).

    Techniques: Migration, Transfection, Negative Control, Incubation