human mmp7 protein  (R&D Systems)

Journal: International Journal of Molecular Sciences

Article Title: Matrix Metalloproteinase 7 Mediates Epithelial–Mesenchymal Transition to Promote Liver Fibrosis Through E-cadherin/β-catenin Pathway in Biliary Atresia

doi: 10.3390/ijms27052209

Figure Lengend Snippet: Increased MMP7 in BA patients correlated with the progression of liver fibrosis. Serum and liver samples of patients with non-BA ( n = 39) disease and BA ( n = 66) were collected. ( A ) Serum MMP7 levels were determined by ELISA. ( B ) Relative MMP7 mRNA levels in livers of BA and non-BA patients. ( C ) Representative images of immunostaining for MMP7 and quantitative analysis of MMP7 staining in liver tissue. Magnification: left panel: ×100; right panel: ×400. ( D ) Representative Masson trichrome staining images of METAVIR fibrosis stages (F1–F4) in BA patients. Scale bar: 400 μm. ( E – G ) Serum MMP7 levels, relative MMP7 mRNA levels and the expression intensity of MMP7 in liver of BA patients with moderate (F0–F2) or advanced liver fibrosis (F3–F4). ( H ) Spearman’s rank correlation analysis of MMP7 expression level with fibrotic marker genes, including COL1A1 , ACTA2 , TGFB1 , and TIMP1 .

Article Snippet: The activation of recombinant human MMP7 protein (R&D Systems, Minneapolis, MN, USA) was achieved through incubation with 4-aminophenylmercuric acetate (APMA; Sigma, St. Louis, MO, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Immunostaining, Staining, Expressing, Marker

Journal: International Journal of Molecular Sciences

Article Title: Matrix Metalloproteinase 7 Mediates Epithelial–Mesenchymal Transition to Promote Liver Fibrosis Through E-cadherin/β-catenin Pathway in Biliary Atresia

doi: 10.3390/ijms27052209

Figure Lengend Snippet: MMP7 gene correlation analysis and pathway enrichment in BA. ( A ) Distribution of correlation coefficients between MMP7 expression and all other genes across two independent BA datasets ( GSE15235 and GSE46960 ). ( B ) GSEA results using gene lists ranked by correlation with MMP7.

Article Snippet: The activation of recombinant human MMP7 protein (R&D Systems, Minneapolis, MN, USA) was achieved through incubation with 4-aminophenylmercuric acetate (APMA; Sigma, St. Louis, MO, USA).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Matrix Metalloproteinase 7 Mediates Epithelial–Mesenchymal Transition to Promote Liver Fibrosis Through E-cadherin/β-catenin Pathway in Biliary Atresia

doi: 10.3390/ijms27052209

Figure Lengend Snippet: EMT scores were associated with MMP7, liver fibrosis and survival in BA patients. Liver samples of patients with non-BA ( n = 39) disease and BA ( n = 66) were collected. ( A ) Representative image of immunostaining for EMT-related protein expression (including E-cadherin, Vimentin, S100A4) in liver biopsies samples. Magnification: left panel : ×100; right panel : ×400. ( B ) Percentage of samples according to the intensity and extension of EMT-related protein immunostaining in cholangiocytes from two groups. ( C ) EMT scores of cholangiocytes in non-BA and BA patients. ( D ) EMT scores in two subgroups divided by MMP7 expression intensity in BA patients. ( E ) Correlation analysis of fibrosis stage EMT scores in BA patients. Spearman’s rank correlation.

Article Snippet: The activation of recombinant human MMP7 protein (R&D Systems, Minneapolis, MN, USA) was achieved through incubation with 4-aminophenylmercuric acetate (APMA; Sigma, St. Louis, MO, USA).

Techniques: Immunostaining, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Matrix Metalloproteinase 7 Mediates Epithelial–Mesenchymal Transition to Promote Liver Fibrosis Through E-cadherin/β-catenin Pathway in Biliary Atresia

doi: 10.3390/ijms27052209

Figure Lengend Snippet: MMP7 promoted EMT in an E-cadherin/β-catenin pathway-dependent manner. ( A ) Protein expression of E-cadherin in HIBEpiCs after treatment with different concentrations of MMP7 for 24 h. ( B ) Level of sE-cadherin in the cell culture supernatant of HIBEpiCs after MMP7 treatment. ( C ) Western blot analysis of β-catenin protein in the nucleus or cytoplasm of HIBEpiCs. ( D ) Immunofluorescence staining shows the subcellular localization of β-catenin in HIBEpiCs. Scale bar:50 μm. ( E ) TOP/FOP-Flash luciferase activity of MMP7-treated cells. ( F ) Effect of the MMP7 and ICG-001 on β-catenin downstream target genes expression (including SNAI1 , SNAI2 , MMP7 ) in HIBEpiCs. ( G , H ) Effect of the MMP7 and ICG-001 on mRNA and protein expression of EMT-related markers (including E-cadherin, Vimentin, S100A4) in HIBEpiCs. ( I ) ELISA analysis of serum level of sE-cadherin in patients. ( J ) Spearman’s rank correlation analysis of serum sE-cadherin and serum MMP7 in BA patients. ( K ) Representative images of immunostaining of β-catenin in cholangiocytes of non-BA and BA patients. Magnification: left panel : ×100; right panel : ×400. ( C – H ) MMP7 concentration: 80 ng/mL; ICG-001 concentration: 25 μM. Data are presented as mean ± SD.

Article Snippet: The activation of recombinant human MMP7 protein (R&D Systems, Minneapolis, MN, USA) was achieved through incubation with 4-aminophenylmercuric acetate (APMA; Sigma, St. Louis, MO, USA).

Techniques: Expressing, Cell Culture, Western Blot, Immunofluorescence, Staining, Luciferase, Activity Assay, Enzyme-linked Immunosorbent Assay, Immunostaining, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Matrix Metalloproteinase 7 Mediates Epithelial–Mesenchymal Transition to Promote Liver Fibrosis Through E-cadherin/β-catenin Pathway in Biliary Atresia

doi: 10.3390/ijms27052209

Figure Lengend Snippet: Fibrosis progression in chronic BA mice was accompanied by increased MMP7 expression and EMT. Liver and serum samples of the control group (i.p. saline) and chronic BA group (i.p. RRV+ anti-Ly6G) were collected on days 14, 21, and 42 postnatally ( n = 5~7 mice in each group). ( A ) Representative micrograph of Masson trichrome staining of the liver from control and chronic BA mice. Quantification of collagen staining in the liver of control and chronic BA mice. Magnification: ×40. ( B , C ) Serum level and hepatic mRNA expression of MMP7 in control and chronic BA mice. ( D ) Representative images of immunostaining for MMP7 expression in liver (Days 42). ( E ) Representative images of immunostaining for EMT-related protein expression (including E-cadherin, Vimentin, S100A4) in liver (Days 42). ( F ) Serum level of sE-cadherin in mice ( n = 5 mice in each group). ( G ) Representative images of immunostaining β-catenin in cholangiocytes of mice. Magnification: left panel : ×100; right panel : ×400. Data are presented as mean ± SD.

Article Snippet: The activation of recombinant human MMP7 protein (R&D Systems, Minneapolis, MN, USA) was achieved through incubation with 4-aminophenylmercuric acetate (APMA; Sigma, St. Louis, MO, USA).

Techniques: Expressing, Control, Saline, Staining, Immunostaining

Journal: International Journal of Molecular Sciences

Article Title: Matrix Metalloproteinase 7 Mediates Epithelial–Mesenchymal Transition to Promote Liver Fibrosis Through E-cadherin/β-catenin Pathway in Biliary Atresia

doi: 10.3390/ijms27052209

Figure Lengend Snippet: MMP7 blockade attenuated liver fibrosis and EMT procession in chronic BA mice. ( A ) Schematic representation of chronic BA mice injected with anti-MMP7 antibody or IgG on days 21–25 and liver specimens collected on day 42 ( n = 5 mice in each group). ( B ) Representative micrographs of Masson trichrome stained liver tissue sections. Quantification of collagen staining in the liver. Magnification: ×40. ( C ) Representative images of immunostaining for EMT-related protein expression (including E-cadherin, Vimentin, S100A4) in the liver (Arrows indicate biliary epithelial cells). ( D ) Serum level of sE-cadherin in mice ( n = 5 mice in each group). ( E ) Representative images of immunostaining β-catenin in cholangiocytes of mice. ( F ) Serum level of MMP7 in mice ( n = 5 mice in each group). ( G ) Hepatic mRNA expression of MMP7 in mice ( n = 5 mice in each group). Magnification: left panel : ×100; right panel : ×400. Data are presented as mean ± SD.

Article Snippet: The activation of recombinant human MMP7 protein (R&D Systems, Minneapolis, MN, USA) was achieved through incubation with 4-aminophenylmercuric acetate (APMA; Sigma, St. Louis, MO, USA).

Techniques: Injection, Staining, Immunostaining, Expressing

Journal: Antibodies (Basel, Switzerland)

Article Title: Characterization of Active MMP9 in Chronic Inflammatory Diseases Using a Novel Anti-MMP9 Antibody.

doi: 10.3390/antib12010009

Figure Lengend Snippet: Figure 1. Activation of MMP9 by distinct proteases in vitro. Western blots for pro-MMP9 (92 kDa, red) and active MMP9 (84 kDa, green) after incubation with active (A) MMP7, (B) MMP12, (C) pepsin A, and (D) plasmin. Below each lane is the MMP9 activity towards gelatin expressed as gelatin fluorescence (arbitrary units) per minute. DQ, dye-quenched; MMP, matrix metalloproteinase.

Article Snippet: Recombinant active MMP7, MMP12, pepsin A, or plasmin (R&D Systems) was titrated against recombinant pro-MMP9 in MMP9 reaction buffer (150 mM NaCl, 10 mM Hepes pH 7.5, 0.05% brij-035, and 5 mM CaCl2).

Techniques: Activation Assay, In Vitro, Western Blot, Incubation, Activity Assay

Journal: Neural Regeneration Research

Article Title: Sequencing analysis of matrix metalloproteinase 7-induced genetic changes in Schwann cells

doi: 10.4103/1673-5374.282263

Figure Lengend Snippet: Heatmap of differentially expressed genes in MMP7-induced Schwann cells. Gene abundances are indicated according to the color bar next to the heatmap. Upregulation is indicated in red, while downregulation is indicated in green. Con: Control; MMP7: matrix metalloproteinase 7.

Article Snippet: Primary SCs were exposed to 10 nM recombinant human MMP7 purified protein (Merck Millipore, Billerica, MA, USA) for 24 hours prior to RNA isolation.

Techniques: Control

Journal: Frontiers in Physiology

Article Title: Peripheral Nerve Injury Induces Dynamic Changes of Tight Junction Components

doi: 10.3389/fphys.2018.01519

Figure Lengend Snippet: Quantitative real-time PCR analysis of tight junction genes in Schwann cells treated with MMPs. (A) MMP7 mRNA expression in Schwann cells transfected with MMP7 siRNA. (B) MMP9 mRNA expression in Schwann cells transfected with MMP9 siRNA. (C–E) The mRNA expression levels of (C) CLDN1, (D) CLDN10, and (E) CLDN22 in Schwann cells transfected with MMP7 siRNA. (F–H) The mRNA expression levels of (F) CLDN1, (G) CLDN10, and (H) CLDN22 in Schwann cells transfected with MMP9 siRNA. The expression levels of tight junction genes were expressed as relative abundance of target genes normalized to MRPL10 with respect to control. Numerical results were shown as mean (SD). ∗ Statistically different from control ( n = 3, paired t -test, P < 0.05).

Article Snippet: Rats in each group received the injection of 10 μl of a 1:1 volume mixture of Matrigel (BD Biosciences, Billerica, MA, United States) with 100 nM human recombinant MMP7 protein (Merck Millipore, Darmstadt, Germany), Matrigel with 100 nM human recombinant MMP9 protein, or Matrigel with 0.9% sodium chloride, respectively, at the crush site immediately after sciatic nerve crush.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Transfection, Control

Journal: Frontiers in Physiology

Article Title: Peripheral Nerve Injury Induces Dynamic Changes of Tight Junction Components

doi: 10.3389/fphys.2018.01519

Figure Lengend Snippet: Quantitative real-time PCR analysis of tight junction genes in the injured sciatic nerve segments injected with MMPs. (A–C) The mRNA expression levels of (A) CLDN1, (B) CLDN10, and (C) CLDN22 at sciatic nerve crush site treated with human recombinant MMP7 protein. (D–F) The mRNA expression levels of (D) CLDN1, (E) CLDN10, and (F) CLDN22 at sciatic nerve crush site treated with human recombinant MMP9 protein. The expression levels of tight junction genes were expressed as relative abundance of target genes normalized to MRPL10 with respect to control. Numerical results were shown as mean (SD). ∗ Statistically different from control ( n = 3, paired t -test, P < 0.05).

Article Snippet: Rats in each group received the injection of 10 μl of a 1:1 volume mixture of Matrigel (BD Biosciences, Billerica, MA, United States) with 100 nM human recombinant MMP7 protein (Merck Millipore, Darmstadt, Germany), Matrigel with 100 nM human recombinant MMP9 protein, or Matrigel with 0.9% sodium chloride, respectively, at the crush site immediately after sciatic nerve crush.

Techniques: Real-time Polymerase Chain Reaction, Injection, Expressing, Recombinant, Control