Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: The Role of Matrix Metalloproteinase-13 (MMP13) in TGFβ/BMP Pathway Regulation of Fibro-Adipogenic Progenitor (FAP) Differentiation.

doi: 10.33594/000000596

Figure Lengend Snippet: Fig. 1. FAPs from MMP13 KO mice have increased spontaneous adi- pogenesis and decreased sponta- neous fibrogenesis compared to FAPs from wildtype mice cultured in a standard medium. A) A typical image of immunostaining for peril- ipin A and αSMA for FAPs from WT and MMP13 KO mice after 2 weeks of culture in standard medium Bottom: negative control of perili- pin A immunofluorescence stain- ing (without primary antibody), negative control of αSMA immu- nofluorescence staining (with- out primary αSMA antibody). B) FAPs from MMP13 KO mice had a significantly higher percentage of perilipin A positive cells when compared to FAPs from WT mice. C) FAPs from MMP13 KO mice had significantly lower αSMA positive cells when compared to FAPs from WT mice. D) Real time PCR showed that FAPs from MMP13 KO mice had significantly higher expression of Adiponectin, but a lower expres- sion of Collagen I compared to FAPs from wildtype mice (* p<0.05).

Article Snippet: MMP13 treatment Recombinant Human MMP13 (R&D systems, 511-MM-010, Minneapolis, MN, USA) were first activated with 1 μM APMA p-Aminophenylmercuric acetate (APMA), (Sigma, Catalog # A-9563, MO, USA) at 37°C for 2 hours.

Techniques: Cell Culture, Immunostaining, Negative Control, Immunofluorescence, Staining, Real-time Polymerase Chain Reaction, Expressing

Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: The Role of Matrix Metalloproteinase-13 (MMP13) in TGFβ/BMP Pathway Regulation of Fibro-Adipogenic Progenitor (FAP) Differentiation.

doi: 10.33594/000000596

Figure Lengend Snippet: Fig. 2. Wildtype FAPs treated with MMP13 inhibitor had sig- nificantly increased adipogen- esis. A) A Typical image of immu- nostaining for FAPs treated with 10μM MMP13 inhibitor and 0.1% DMSO in standard medium for 2 weeks. B) Wildtype FAPs treated with MMP13 inhibitor had a sig- nificantly higher percentage of perilipin A(+) cells compared to those treated with DMSO. C) FAPs treated with MMP13 inhibitor had a significantly reduced number of αSMA(+) cells compared to those treated with DMSO. D) Real time PCR showed that FAPs treated with the MMP13 inhibitor had a sig- nificantly increased expression of Adiponectin, PPARγ, C/EBPA and deceased expression of αSMA and Collagen 1a (* p<0.05).

Article Snippet: MMP13 treatment Recombinant Human MMP13 (R&D systems, 511-MM-010, Minneapolis, MN, USA) were first activated with 1 μM APMA p-Aminophenylmercuric acetate (APMA), (Sigma, Catalog # A-9563, MO, USA) at 37°C for 2 hours.

Techniques: Real-time Polymerase Chain Reaction, Expressing

Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: The Role of Matrix Metalloproteinase-13 (MMP13) in TGFβ/BMP Pathway Regulation of Fibro-Adipogenic Progenitor (FAP) Differentiation.

doi: 10.33594/000000596

Figure Lengend Snippet: Fig. 3. MMP13 treatment inhibits FAP adipogenesis and promotes FAP fibrogenesis. A) A typical im- age of immunostaining for FAPs treated with 100 ng/ml MMP13 and 0.1% DMSO in standard me- dium for 2 weeks. B) FAPs treated with MMP13 have a significantly reduced the number of perili- pin (+) cells compared to DMSO. C) FAPs treated with MMP13 have a significantly increased the num- ber of αSMA (+) cells compared to DMSO. D) Real time PCR results of FAPs treated with MMP13 have a significantly higher expression of αSMA and collagen I and decreased expression of Adiponectin and PPARγ (* p<0.05).

Article Snippet: MMP13 treatment Recombinant Human MMP13 (R&D systems, 511-MM-010, Minneapolis, MN, USA) were first activated with 1 μM APMA p-Aminophenylmercuric acetate (APMA), (Sigma, Catalog # A-9563, MO, USA) at 37°C for 2 hours.

Techniques: Immunostaining, Real-time Polymerase Chain Reaction, Expressing

Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: The Role of Matrix Metalloproteinase-13 (MMP13) in TGFβ/BMP Pathway Regulation of Fibro-Adipogenic Progenitor (FAP) Differentiation.

doi: 10.33594/000000596

Figure Lengend Snippet: Fig. 4. A) The typical images of FAPs from wildtype and MMP13 KO mice treated with TGFβ-1, TGFβ in- hibitor, or 0.1% DMSO in a standard medium for 2 weeks. B) TGFβ-1 significantly decreased the number of perilipin (+) cells and the TGFβ inhibitor significantly increased the number of perilipin (+) cells in FAP WT mice, but not in FAPs from MMP13 KO mice. C) TGFβ-1 significantly increased the number of αSMA (+) cells and the TGFβ inhibitor significantly decreased the number of αSMA (+) cells in FAPs from WT mice, but not in FAPs from MMP13 KO mice. D) & E) Real time PCR results showed that TGFβ-1 significantly decreased the expression of adipogenesis-related genes and increased fibrogenesis-related gene expression of FAPs in WT mice, while the TGFβ inhibitor had an opposite effect. However, the effect of the TGFβ-1 and TGFβ inhibitors had no effect on their expression in FAPs from MMP13 KO mice ( * p<0.05). F) Exogenous MMP13 added to the TGFβ inhibitor treatment group in both WT mice and MMP13 KO mice. Exogenous MMP13 added to the MMP-13 KO mice as a control. G) Quantification of the percentage of the number of Perilipin A(+) cells and αSMA (+) cells out of total number of cells.

Article Snippet: MMP13 treatment Recombinant Human MMP13 (R&D systems, 511-MM-010, Minneapolis, MN, USA) were first activated with 1 μM APMA p-Aminophenylmercuric acetate (APMA), (Sigma, Catalog # A-9563, MO, USA) at 37°C for 2 hours.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Gene Expression, Control

Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: The Role of Matrix Metalloproteinase-13 (MMP13) in TGFβ/BMP Pathway Regulation of Fibro-Adipogenic Progenitor (FAP) Differentiation.

doi: 10.33594/000000596

Figure Lengend Snippet: Fig. 5. A) Typical images of FAPs from WT and MMP13 KO mice treated with BMP-7, BMP inhibitor, or 0.1% DMSO in standard medium for 2 weeks. B) BMP-7 significantly increased the number of perilipin (+) cells and the BMP inhibitor significantly decreased the number of perilipin (+) cells in FAPs in WT mice, but not in FAPs from MMP13 KO mice. C) BMP-7 significantly decreased αSMA (+) cells and the BMP inhibitor sig- nificantly increased the number of αSMA (+) cells in FAPs in WT mice, but not in FAPs from MMP13 KO mice. D) & E) Real time PCR results showed that BMP-7 significantly increased the expression of adipogenesis- related genes and decreased fibrogenesis-related gene expression of FAPs in WT mice, while BMP inhibitor had an opposite effect. Neither BMP-7 nor BMP inhibitor showed an effect on adipogenesis-related and fibrogenesis-related gene expression in FAPs from MMP13 KO mice ( * p<0.05). F) Exogenous MMP13 added to the BMP inhibitor treatment group in both WT mice and MMP13 KO mice. Exogenous MMP13 was added to the MMP13 KO mice as a control. G) Quantification of Perilipin A and αSMA.

Article Snippet: MMP13 treatment Recombinant Human MMP13 (R&D systems, 511-MM-010, Minneapolis, MN, USA) were first activated with 1 μM APMA p-Aminophenylmercuric acetate (APMA), (Sigma, Catalog # A-9563, MO, USA) at 37°C for 2 hours.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Gene Expression, Control

Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: The Role of Matrix Metalloproteinase-13 (MMP13) in TGFβ/BMP Pathway Regulation of Fibro-Adipogenic Progenitor (FAP) Differentiation.

doi: 10.33594/000000596

Figure Lengend Snippet: Fig. 6. A) Typical images of FAPs from WT mice treated with the MMP13 inhibitor (or DMSO) in combina- tion of TGFβ-1 and TGFβ inhibitor in standard medium for 2 weeks. B) TGFβ-1 significantly decreased the number of perilipin A(+) cells and TGFβ inhibitor significantly increased the number of perilipin A (+) cells in FAPs without the MMP13 inhibitor. However, MMP13 inhibitor blocked the effect of TGFβ-1 and TGFβ inhibitors. C) TGFβ-1 significantly increased the numbers of αSMA (+) cells and the TGFβ inhibitor signifi- cantly decreased the number of αSMA (+) cells in FAPs without the MMP13 inhibitor. However, the MMP13 inhibitor blocked the effects of the TGFβ-1 and TGFβ inhibitor. D) & E) Real time PCR results showed that TGFβ-1 significantly decreased adipogenesis-related gene expression and increased fibrogenesis-related gene expression of FAPs without the MMP13 inhibitor, but not in combination with the MMP13 inhibitor (* p<0.05).

Article Snippet: MMP13 treatment Recombinant Human MMP13 (R&D systems, 511-MM-010, Minneapolis, MN, USA) were first activated with 1 μM APMA p-Aminophenylmercuric acetate (APMA), (Sigma, Catalog # A-9563, MO, USA) at 37°C for 2 hours.

Techniques: Real-time Polymerase Chain Reaction, Gene Expression

Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: The Role of Matrix Metalloproteinase-13 (MMP13) in TGFβ/BMP Pathway Regulation of Fibro-Adipogenic Progenitor (FAP) Differentiation.

doi: 10.33594/000000596

Figure Lengend Snippet: Fig. 7. A) Typical images of FAPs from WT mice treated with an MMP13 inhibitor (or DMSO) in combination with BMP-7 and the BMP inhibitor in standard medium for 2 weeks. B) BMP-7 significantly increased the number of perilipin (+) cells and BMP inhibitor significantly decreased the number of perilipin (+) cells in FAPs without the MMP13 inhibitor, but not in combination with the MMP13 inhibitor. C) BMP-7 significantly decreased the number of αSMA (+) cells and the BMP inhibitor significantly increased the number of αSMA (+) cells in FAPs from WT mice, but not in FAPs from MMP13 KO mice. D) & E) Real time PCR results showed that BMP significantly increased the expression of adipogenesis-related genes and decreased the expression of fibrogenesis-related in FAPs from WT mice, while the BMP inhibitor had an opposite effect. However, the effect of BMP-7 and BMP inhibitor was abolished in FAPs from MMP13 KO mice (* p<0.05).

Article Snippet: MMP13 treatment Recombinant Human MMP13 (R&D systems, 511-MM-010, Minneapolis, MN, USA) were first activated with 1 μM APMA p-Aminophenylmercuric acetate (APMA), (Sigma, Catalog # A-9563, MO, USA) at 37°C for 2 hours.

Techniques: Real-time Polymerase Chain Reaction, Expressing