Fig 2: Knockdown of ACVR2A suppresses Con A-induced activation of hepatic stellate cells in vivo. (A) Relative mRNA and (B) protein expression levels of ACTA2, COL1A1 and COL4A1 were determined using reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. (C) Immunohistochemistry was performed to detect the protein expression levels of α-SMA, collagen I and IV. Orange arrowheads, representative α-SMA-positive cells; pink arrowheads, representative collagen I-positive cells; green arrowheads, representative collagen IV-positive cells. Scale bars, 50 μm. (D) Protein expression levels of p-Smad2 and total Smad2 in liver tissues were detected using western blot analysis. Data are expressed as the means ± standard error (n=5/group). α-SMA, α-smooth muscle actin; ACVR2A, activin A receptor type 2A; Con A, concanavalin A; ; NC, negative control; p-, phosphorylated; shRNA, short hairpin RNA.

Fig 3: Glycyrrhizic acid inhibited EMT and activation of TGF-β signaling in BLM-induced pulmonary fibrosis. (A) Immunofluorescence staining of E-cadherin (E-cad) in the lungs. The sections were photographed at 400× magnification (scale bars = 50 μm). A small window is enlarged by five times to present the subcellular location of E-cad. (B) Immunoblotting for the epithelial marker (E-cad), myofibroblasts maker (α-SMA), and ECM molecules (Fibronectin and Vimenin) in the lung tissues. (C) Densitometric analysis of proteins of interest in the immunoblots using β-actin as the internal reference. Values are expressed as the mean ± standard deviation (n = 8 per group). Compared with the Control group, **p < 0.01, ***p < 0.001; Compared with the BLM group, #p < 0.05, ##p < 0.01, ###p < 0.001. (D) Immunoblotting for TGF-β1 and its downstream signaling molecules (p-Smad2 and p-Smad3) in the lung tissues. (E) Densitometric analysis of target proteins in (D) using β-actin as the internal reference. Values are expressed as the mean ± standard deviation (n = 8 per group). Compared with the Control group, ***p < 0.001; Compared with the BLM group, ##p < 0.01, ###p < 0.001.

Fig 4: The expression of phosphorylated TGF-β1, TGF-βR1 and Smad2 in rabbit endometrium. a The expression of TGF-β1, TGF-βR1, and Smad2 and their phosphorylated forms detected by Western blot at different time points. The protein expressions of b TGF-β1, c TGF-βR1, and d Smad2 were quantified in relative to that of GAPDH. e The percentage of p-TGF-β1 in the total TGF-β1 protein. f The percentage of p- TGF-βR1 in the total TGF-βR1 protein. g The percentage of p-Smad2 in the total Smad2 protein. *P < 0.05, compared to the Sham operation group; #P < 0.05 compared to the model group; ΔP < 0.05, comparison between the expressions at 2 weeks and 4 weeks

Fig 5: Effects of CAV1 on C2C12 cell differentiation. (A). Change in CAV1, MYH2, and MYOG expression levels at different stages of C2C12 cell differentiation. (B). Grayscale scan of CAV1 in Figure A. (C). Grayscale scan of MYH2 in Figure A. (D). Grayscale scan of MYOG in Figure A. (E). Immunofluorescence was used to measure the changes in CAV1 expression levels and the morphology of myotubes at various stages of C2C12 differentiation. CAV1 was labeled green using FITC and the nucleus blue using DAPI. (F). Laser confocal microscopy demonstrating the location of CAV1 expression, stained red using RBFITC, and nuclei blue using DAPI. (G). Changes in CAV1, MYH2, and MYOG expression levels after siRNA transfection for 48 h and 72 h. (H). Grayscale scan of CAV1 in Figure G. (I). Grayscale scan of MYH2 in Figure G. (J). Grayscale scan of MYOG in Figure G. (K). Changes in the fusion rate of C2C12 cells in mice after CAV1 inhibition by immunofluorescence detection. (L). Myotubule fusion rate in Figure K. (M). Change in CAV1, P-Smad2, and P-Smad4 expression after siRNA fragment S1 transfection of CAV1. (N). Grayscale scan of CAV1 in Figure M. (O). Grayscale scan of P-Smad2 in Figure M. (P). Grayscale scan of P-Smad4 in Figure M. (** p values < 0.01, * p values < 0.05) (n = 3).