recombinant tnfα standards  (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