Fig. 5
TGF-β1-miR29a-Fstl1 Regulatory Circuit Mediates Fibrosis. a The livers of Fstl1+/− mice were harvested for the analyses. The expression of miR29a was assessed by qRT-PCR (n = 3 per group). b Fstl1 siRNA (40 nM) transfection was performed on LX-2 cells using Lipofectamine RNAimax (n = 3 per group). The expression of miR29a was assessed by qRT-PCR (n = 3 per group). c BABL/c mice were subjected to CCl4 along with Fstl1-neutralizing antibody 22B6 or isotype control IgG1 (25 μg/mouse) twice a week, and livers were harvested 4 weeks later for the analyses. The expression of miR29a was assessed by qRT-PCR (n = 6 per group). d LX-2 cells were treated with 22B6 or isotype control IgG1(2 μg/ml). The expression of miR29a was assessed by qRT-PCR (n = 3 per group). e MiR29a mimics (100 nM) were transiently transfected into LX-2 cells for 48 h. The expression of Fstl1, Col1, α-SMA mRNA and miR29a were assessed by qRT-PCR (n = 3 per group). f MiR29a inhibitors (200 nM) were transiently transfected into LX-2 cells for 48 h. The expression of Fstl1, Col1, α-SMA mRNA and miR29a were assessed by qRT-PCR (n = 3 per group). g-h Protein expression levels of α-SMA and Fstl1 in cell extracts were assessed by Western blot. Band intensity was quantified using Image J software and expressed as relative intensity compared with control. The ratio of Fstl1 and α-SMA were subjected to GAPDH (n = 2 per group). Throughout, data represent means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001