recombinant human mmp  (R&D Systems)

Journal: The FASEB Journal

Article Title: CBP /β‐Catenin Signaling in Hepatocytes Plays Pivotal Roles in MMP ‐7‐Mediated Liver Fibrosis in a Metabolic Dysfunction‐Associated Steatohepatitis Mouse Model

doi: 10.1096/fj.202504510R

Figure Lengend Snippet: Intrahepatic expression of MMP‐7 is markedly reduced in CBP hep‐KO mice. Male CBP hep‐KO mice (8–10 weeks old, n = 3) and P300 hep‐KO mice ( n = 3) were fed with a CDAHFD for 16 weeks. (A) Differentially expressed genes (DEGs). The volcano plot shows the results for the DEGs. (B) GOBP analysis of the top 10 downregulated genes in the livers of CBP hep‐KO mice. (C) Clustergrams of PCR array analyses of WNT/β‐catenin‐related genes (WNT/β‐catenin signaling pathway). Green indicates low expression levels, while red indicates increased expression levels. (D) MMP‐7 mRNA expression in the liver, as determined by RT‐qPCR ( n = 9, 8, and 13 per group, respectively). The results shown are representative of at least three independent experiments. Data are presented as mean ± SD. * p < 0.05; ** p < 0.01; *** p < 0.005; **** p < 0.0001, by one‐way ANOVA.

Article Snippet: The LX‐2 human hepatic stellate cell line (SCC064; Sigma‐Aldrich) was treated with recombinant human MMP‐7 (R&D Systems, 907‐MP) at concentrations ranging from 0.05–0.5 nM for 24 h prior to RNA extraction.

Techniques: Expressing, Quantitative RT-PCR

Journal: The FASEB Journal

Article Title: CBP /β‐Catenin Signaling in Hepatocytes Plays Pivotal Roles in MMP ‐7‐Mediated Liver Fibrosis in a Metabolic Dysfunction‐Associated Steatohepatitis Mouse Model

doi: 10.1096/fj.202504510R

Figure Lengend Snippet: Effects of MMP‐7 on hepatic stellate cells. (A) Schematic of the treatment protocol. HepG2 cells were treated with palmitate (200 μM, 500 μM) for 24 h. The cells were then harvested, RNA was extracted, and MMP‐7 expression was analyzed using RT‐PCR. (B) Schematic representation of the treatment protocol. After culturing LX‐2 cells for 24 h, human recombinant MMP‐7 was added at concentrations ranging from 0.05 to 0.5 nM. Cells were harvested after 24 h, RNA was extracted, and the expression of αSMA and Col1a1 was analyzed by rt‐PCR. The results are representative of at least three independent experiments. Data represent the mean ± SD. * p < 0.05; ** p < 0.01; *** p < 0.005; **** p < 0.0001, by one‐way ANOVA.

Article Snippet: The LX‐2 human hepatic stellate cell line (SCC064; Sigma‐Aldrich) was treated with recombinant human MMP‐7 (R&D Systems, 907‐MP) at concentrations ranging from 0.05–0.5 nM for 24 h prior to RNA extraction.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Recombinant

Journal: The FASEB Journal

Article Title: CBP /β‐Catenin Signaling in Hepatocytes Plays Pivotal Roles in MMP ‐7‐Mediated Liver Fibrosis in a Metabolic Dysfunction‐Associated Steatohepatitis Mouse Model

doi: 10.1096/fj.202504510R

Figure Lengend Snippet: Serum MMP‐7 levels are increased in patients with MASH. (A) MMP‐7 concentrations were measured in serum samples from patients with MASH ( n = 54) and healthy volunteers ( n = 70). (B) MMP‐7 levels were measured in serum samples from patients with MASLD or MASH who underwent liver biopsy and had their fibrosis grade determined. F0 ( n = 16), F1‐2 ( n = 20), and F3‐4 ( n = 22). (C) Representative immunohistochemical images of CBP and P300 in normal ( n = 5) and NASH ( n = 6) liver biopsy specimens. Increased nuclear CBP staining was observed in hepatocytes from NASH livers. The number of cells positive for CBP and P300 in the liver was determined. For each sample, the number of positive cells was counted in five fields at 20× magnification. (D) Quantification of CBP‐positive and P300‐positive hepatocytes in liver biopsy specimens. The number of positive cells was counted in five fields at 20× magnification.

Article Snippet: The LX‐2 human hepatic stellate cell line (SCC064; Sigma‐Aldrich) was treated with recombinant human MMP‐7 (R&D Systems, 907‐MP) at concentrations ranging from 0.05–0.5 nM for 24 h prior to RNA extraction.

Techniques: Immunohistochemical staining, Staining

Journal: Scientific Reports

Article Title: Nisin bacteriocin blocks T. denticola -triggered MMP2 activation and pathogen internalization via TLR2

doi: 10.1038/s41598-026-43673-8

Figure Lengend Snippet: Nisin inhibits T. denticola -induced-MMP2 activation in PDL cells. ( a ) Representative gelatin zymogram showing MMP2 levels in PDL cells exposed to wild-type T. denticola (35405) (50 MOI) for 2 h, washed three times with PBS, followed by treatment with nisin (100 µg/mL) for 24 h. Conditioned media were analyzed by gelatin zymography. Quantitative analysis of band intensities for total MMP2 ( b ), Pro-MMP2 (c), active MMP2 (d) and dentilisin/PrtP (e). Data are representative of at least three independent experiments. Intergroup differences were analyzed by the analysis of variance (ANOVA) and Tukey’s post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. The uncropped gel image is presented in Supplementary Information as Fig. 1a.

Article Snippet: For in vitro MMP2 activation, different concentrations of purified dentilisin/PrtP (20, 40 or 80 ng) , and pro-MMP2 (20 ng) (Cat #902-MP-010; R&D Systems, Minneapolis, MN) diluted in reaction buffer (TCNB, 50 mM Tris, 10 mM CaCl 2 , 150 mM NaCl, 0.05% (w/v) Brij 35, pH 7.5) were mixed and incubated for 30 min at room temperature, then mixed with 4X sample buffer (Cat #1610747, Bio-Rad, Hercules, CA) and loaded onto 10% gelatin-containing SDS-PAGE gels (Cat #ZY00100BOX, Thermo Fisher scientific, Novex TM 10% zymogram plus (Gelatin) protein gels).

Techniques: Activation Assay, Zymography

Journal: Scientific Reports

Article Title: Nisin bacteriocin blocks T. denticola -triggered MMP2 activation and pathogen internalization via TLR2

doi: 10.1038/s41598-026-43673-8

Figure Lengend Snippet: T. denticola purified dentilisin/PrtP directly activates recombinant MMP2 in vitro, and nisin attenuates this activation. ( a ) Representative gelatin zymogram of recombinant Pro-MMP2 (20 ng) incubated with increasing concentrations of purified dentilisin/PrtP (20, 40, or 80 ng) in the presence or absence of nisin (20, 40, or 80 µg). Quantitative analysis of band intensities for total MMP2 ( b ), Pro-MMP2 ( c ), active MMP2 ( d ) and dentilisin/PrtP ( e ). Data are representative of at least three independent experiments. Intergroup differences were analyzed by the analysis of variance (ANOVA) and Tukey’s post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. The uncropped gel image is presented in Supplementary Information as Fig. 2a.

Article Snippet: For in vitro MMP2 activation, different concentrations of purified dentilisin/PrtP (20, 40 or 80 ng) , and pro-MMP2 (20 ng) (Cat #902-MP-010; R&D Systems, Minneapolis, MN) diluted in reaction buffer (TCNB, 50 mM Tris, 10 mM CaCl 2 , 150 mM NaCl, 0.05% (w/v) Brij 35, pH 7.5) were mixed and incubated for 30 min at room temperature, then mixed with 4X sample buffer (Cat #1610747, Bio-Rad, Hercules, CA) and loaded onto 10% gelatin-containing SDS-PAGE gels (Cat #ZY00100BOX, Thermo Fisher scientific, Novex TM 10% zymogram plus (Gelatin) protein gels).

Techniques: Purification, Recombinant, In Vitro, Activation Assay, Incubation

Journal: Scientific Reports

Article Title: Nisin bacteriocin blocks T. denticola -triggered MMP2 activation and pathogen internalization via TLR2

doi: 10.1038/s41598-026-43673-8

Figure Lengend Snippet: Dentilisin/PrtP-deficient T. denticola does not activate MMP2 and is unaffected by nisin in PDL cells. ( a ) Representative gelatin zymogram showing MMP2 levels in PDL cells exposed to purified dentilisin/PrtP (50 ng/ml), wild-type T. denticola (WT) (35405) (50 MOI) or mutant T. denticola (MT) (50 MOI) for 2 h and treated with nisin (100 µg/ml) for 24 h. Conditioned media were analyzed by gelatin zymography. Quantitative analysis of band intensities for total MMP2 ( b ), Pro-MMP2 ( c ), active MMP2 ( d ) and dentilisin/PrtP ( e ). Data are representative of at least three independent experiments. Intergroup differences were analyzed by the analysis of variance (ANOVA) and Tukey’s post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. The uncropped gel image is presented in Supplementary Information as Fig. 3a.

Article Snippet: For in vitro MMP2 activation, different concentrations of purified dentilisin/PrtP (20, 40 or 80 ng) , and pro-MMP2 (20 ng) (Cat #902-MP-010; R&D Systems, Minneapolis, MN) diluted in reaction buffer (TCNB, 50 mM Tris, 10 mM CaCl 2 , 150 mM NaCl, 0.05% (w/v) Brij 35, pH 7.5) were mixed and incubated for 30 min at room temperature, then mixed with 4X sample buffer (Cat #1610747, Bio-Rad, Hercules, CA) and loaded onto 10% gelatin-containing SDS-PAGE gels (Cat #ZY00100BOX, Thermo Fisher scientific, Novex TM 10% zymogram plus (Gelatin) protein gels).

Techniques: Purification, Mutagenesis, Zymography

Journal: Scientific Reports

Article Title: Nisin bacteriocin blocks T. denticola -triggered MMP2 activation and pathogen internalization via TLR2

doi: 10.1038/s41598-026-43673-8

Figure Lengend Snippet: T. denticola -triggered activation of MMP2 and nisin effects are mediated via TLR2 in PDL cells. PDL cells were transduced with TLR2 shRNA or scramble shRNA lentiviral particles in serum-free media, then selected in puromycin. ( a ) Immunoblot showing TLR2 protein levels in cells transduced with scramble shRNA or TLR2 shRNA lentiviral particles. ( b ) Representative gelatin zymogram showing MMP2 levels in PDL cells exposed to wild-type T. denticola (35405) (50 MOI) for 2 h, washed three times with PBS, treated with gentamicin (50 µg/ml) for 1 h, washed three times again with PBS, and then treated with nisin (100 µg/ml) for 24 h. Conditioned media were analyzed by gelatin zymography. Quantitative analysis of band intensities for total MMP2 ( c ), Pro-MMP2 ( d ), active MMP2 ( e ) and dentilisin/PrtP ( f ). Data are representative of at least three independent experiments. Intergroup differences were analyzed by the analysis of variance (ANOVA) and Tukey’s post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. The uncropped blot and gel images are presented in Supplementary Information as Figs. 4a and b.

Article Snippet: For in vitro MMP2 activation, different concentrations of purified dentilisin/PrtP (20, 40 or 80 ng) , and pro-MMP2 (20 ng) (Cat #902-MP-010; R&D Systems, Minneapolis, MN) diluted in reaction buffer (TCNB, 50 mM Tris, 10 mM CaCl 2 , 150 mM NaCl, 0.05% (w/v) Brij 35, pH 7.5) were mixed and incubated for 30 min at room temperature, then mixed with 4X sample buffer (Cat #1610747, Bio-Rad, Hercules, CA) and loaded onto 10% gelatin-containing SDS-PAGE gels (Cat #ZY00100BOX, Thermo Fisher scientific, Novex TM 10% zymogram plus (Gelatin) protein gels).

Techniques: Activation Assay, Transduction, shRNA, Western Blot, Zymography