Journal: Molecular Medicine Reports

Article Title: Cathepsin S regulates renal fibrosis in mouse models of mild and severe hydronephrosis

doi: 10.3892/mmr.2019.10230

Figure Lengend Snippet: CTSS inhibitor treatment decreased the expression of CTSS in mouse kidneys, whereas TGF-β1 treatment increased the expression of CTSS in TCMK-1 cells. (A and D) CTSS measured by immunohistochemistry (magnification, ×400) in mice treated with LY3000328 or not; (B and C) CTSS measured by WB in mice treated with LY3000328 or not; (E) CTSS measured by qPCR in mice treated with LY3000328 or not; (F and G) CTSS measured by WB in TCMK-1 cells stimulated by different concentrations of TGF-β1; (H) CTSS measured by qPCR in TCMK-1 cells stimulated by different concentrations of TGF-β1; (I and J) CTSS measured by WB in TCMK-1 cells treated by siRNA-CTSS; (K) CTSS measured by qPCR in TCMK-1 cells treated by siRNA-CTSS. The data are shown as the mean ± standard deviation; # P<0.05 vs. the N group, & P<0.05 vs. the M group, *P<0.05 vs. the S group. Grouping: The N groups (N-no inhibitors, Ni-inhibitors), no hydronephrosis; the M groups (M-no inhibitors, Mi-inhibitors), mild hydronephrosis; and the S groups (S-no inhibitors, Si-inhibitors), severe hydronephrosis. CTSS, cathepsin S; TGF, transforming growth factor; WB, western blotting; qPCR, quantitative PCR.

Article Snippet: Endogenous peroxidase was removed using 3% hydrogen peroxide and the following primary antibodies were added: α-Smooth muscle actin [α-SMA; 1:200; cat. no. 19245T; Cell Signaling Technology, Inc. (CST)], E-cadherin (1:400; cat. no. 3195T; CST), transforming growth factor β1 (TGF-β1; 1:100; cat. no. ab92486; Abcam), CTSS (1:50; cat. no. Sc271619; Santa Cruz Biotechnology, Inc.), collagen-I (1:100; cat. no. ab34710; Abcam) and fibronectin (FN; 1:200; cat. no. ab2413; Abcam).

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

Journal: Molecular Medicine Reports

Article Title: Cathepsin S regulates renal fibrosis in mouse models of mild and severe hydronephrosis

doi: 10.3892/mmr.2019.10230

Figure Lengend Snippet: Reducing the expression of CTSS can increase the deposition of ECM. (A, B, D and E) COL-1 and FN measured by immunohistochemistry (magnification, ×400) in mice treated with LY3000328 or not; (C, F and G) COL-1 and FN measured by WB in mice treated with LY3000328 or not; (H and I) COL-1 and FN measured by qPCR in mice treated with LY3000328 or not; (J-L) COL-1 and FN measured by WB in TCMK-1 cells treated with TGF-β or not; (M and N) COL-1 and FN measured by qPCR in TCMK-1 cells treated with TGF-β or not. The data are shown as the mean ± standard deviation; # P<0.05 vs. the N group or corresponding control (CON), & P<0.05 vs. the M group or corresponding scramble, *P<0.05 vs. the S group or corresponding control (CON). Grouping: The N groups (N-no inhibitors, Ni-inhibitors), no hydronephrosis; the M groups (M-no inhibitors, Mi-inhibitors), mild hydronephrosis; and the S groups (S-no inhibitors, Si-inhibitors), severe hydronephrosis. CTSS, cathepsin S; ECM, extracellular matrix; WB, western blotting; qPCR, quantitative PCR.

Article Snippet: Endogenous peroxidase was removed using 3% hydrogen peroxide and the following primary antibodies were added: α-Smooth muscle actin [α-SMA; 1:200; cat. no. 19245T; Cell Signaling Technology, Inc. (CST)], E-cadherin (1:400; cat. no. 3195T; CST), transforming growth factor β1 (TGF-β1; 1:100; cat. no. ab92486; Abcam), CTSS (1:50; cat. no. Sc271619; Santa Cruz Biotechnology, Inc.), collagen-I (1:100; cat. no. ab34710; Abcam) and fibronectin (FN; 1:200; cat. no. ab2413; Abcam).

Techniques: Expressing, Immunohistochemistry, Standard Deviation, Control, Western Blot, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Cathepsin S regulates renal fibrosis in mouse models of mild and severe hydronephrosis

doi: 10.3892/mmr.2019.10230

Figure Lengend Snippet: The expression of TGF-β signaling pathway-related factors changes after interference with the expression of CTSS. (A, C-G) Smad2/3, pSmad2/3 and TGF-β measured by WB in mice treated with LY3000328 or not; (B, I-L) Smad2/3, pSmad2/3 and TGF-β measured by WB in TCMK-1 cells treated with TGF-β or not; (H) TGF-β measured by qPCR in mice treated with LY3000328 or not. The data are shown as the mean ± standard deviation; # P<0.05 vs. the N group or corresponding control (CON), & P<0.05 vs. the M group or corresponding scramble, *P<0.05 vs. the S group or corresponding control (CON). Grouping: The N groups (N-no inhibitors, Ni-inhibitors), no hydronephrosis; the M groups (M-no inhibitors, Mi-inhibitors), mild hydronephrosis; and the S groups (S-no inhibitors, Si-inhibitors), severe hydronephrosis. TGF, transforming growth factor; CTSS, cathepsin S; WB, western blotting; qPCR, quantitative PCR.

Article Snippet: Endogenous peroxidase was removed using 3% hydrogen peroxide and the following primary antibodies were added: α-Smooth muscle actin [α-SMA; 1:200; cat. no. 19245T; Cell Signaling Technology, Inc. (CST)], E-cadherin (1:400; cat. no. 3195T; CST), transforming growth factor β1 (TGF-β1; 1:100; cat. no. ab92486; Abcam), CTSS (1:50; cat. no. Sc271619; Santa Cruz Biotechnology, Inc.), collagen-I (1:100; cat. no. ab34710; Abcam) and fibronectin (FN; 1:200; cat. no. ab2413; Abcam).

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

Journal: Molecular Medicine Reports

Article Title: Cathepsin S regulates renal fibrosis in mouse models of mild and severe hydronephrosis

doi: 10.3892/mmr.2019.10230

Figure Lengend Snippet: Reducing the expression of CTSS can exacerbate EMT. (A, B, D and E) α-SMA and E-cadherin measured by immunohistochemistry (magnification, ×400) in mice treated with LY3000328 or not; (C, F and G) α-SMA and E-cadherin measured by WB in mice treated with LY3000328 or not; (H and I) α-SMA and E-cadherin measured by qPCR in mice treated with LY3000328 or not; (J-L) α-SMA and E-cadherin measured by WB in TCMK-1 cells treated with TGF-β or not; (M and N) α-SMA and E-cadherin measured by qPCR in TCMK-1 cells treated with TGF-β or not. The data are shown as the mean ± standard deviation; # P<0.05 vs. the N group or corresponding control (CON), & P<0.05 vs. the M group or corresponding scramble, *P<0.05 vs. the S group or corresponding control (CON). Grouping: The N groups (N-no inhibitors, Ni-inhibitors), no hydronephrosis; the M groups (M-no inhibitors, Mi-inhibitors), mild hydronephrosis; and the S groups (S-no inhibitors, Si-inhibitors), severe hydronephrosis. CTSS, cathepsin S; EMT, epithelial-mesenchymal transition; WB, western blotting; qPCR, quantitative PCR; TGF, transforming growth factor.

Article Snippet: Endogenous peroxidase was removed using 3% hydrogen peroxide and the following primary antibodies were added: α-Smooth muscle actin [α-SMA; 1:200; cat. no. 19245T; Cell Signaling Technology, Inc. (CST)], E-cadherin (1:400; cat. no. 3195T; CST), transforming growth factor β1 (TGF-β1; 1:100; cat. no. ab92486; Abcam), CTSS (1:50; cat. no. Sc271619; Santa Cruz Biotechnology, Inc.), collagen-I (1:100; cat. no. ab34710; Abcam) and fibronectin (FN; 1:200; cat. no. ab2413; Abcam).

Techniques: Expressing, Immunohistochemistry, Standard Deviation, Control, Western Blot, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Cathepsin S regulates renal fibrosis in mouse models of mild and severe hydronephrosis

doi: 10.3892/mmr.2019.10230

Figure Lengend Snippet: Interfering with the expression of CTSS aggravates renal fibrosis. (A) Masson staining (magnification, ×200); (B) Quantitative analysis of the degree of fibrosis. The blue area represents the deposition of collagen. The data are shown as the mean ± standard deviation; # P<0.05 vs. the N group, & P<0.05 vs. the M group, *P<0.05 vs. the S group. Grouping: The N groups (N-no inhibitors, Ni-inhibitors), no hydronephrosis; the M groups (M-no inhibitors, Mi-inhibitors), mild hydronephrosis; and the S groups (S-no inhibitors, Si-inhibitors), severe hydronephrosis. CTSS, cathepsin S.

Article Snippet: Endogenous peroxidase was removed using 3% hydrogen peroxide and the following primary antibodies were added: α-Smooth muscle actin [α-SMA; 1:200; cat. no. 19245T; Cell Signaling Technology, Inc. (CST)], E-cadherin (1:400; cat. no. 3195T; CST), transforming growth factor β1 (TGF-β1; 1:100; cat. no. ab92486; Abcam), CTSS (1:50; cat. no. Sc271619; Santa Cruz Biotechnology, Inc.), collagen-I (1:100; cat. no. ab34710; Abcam) and fibronectin (FN; 1:200; cat. no. ab2413; Abcam).

Techniques: Expressing, Staining, Standard Deviation