Figure 7.

miR-101 controls α-SMA and collagen expression in lung fibroblasts. a and b, effect of miR-101 overexpression on TGF-β1-induced α-SMA and collagen protein and mRNA expression. LL29 cells were treated with lentiviral miR-101 or the virus control (vc) at an m.o.i. of 50 for 48 h. Then, the cells were stimulated with 5 ng/ml TGF-β1 for 48 h. Western blotting and real-time PCR were performed to determine the protein and mRNA expression levels. The expression levels were relative to GAPDH. n = 3 cell preparations. c and d, effect of miR-101 knockdown on the protein and mRNA expression of α-SMA, COL1A1, and COL3A1. CCD-8Lu fibroblasts were treated with lentiviral anti-miR-101 or the virus control at an m.o.i. of 50 for 72 h, and protein and mRNA expression was determined by Western blotting and real-time PCR. The expression levels were relative to GAPDH from two cell preparations, each performed in duplicate. e, miR-101 suppresses TGFβ-1-induced collagen gel contraction in LL29 cells. f, LL29 cells were treated with lentiviral miR-101 or the virus control at an m.o.i. of 50 for 48 h. Then, the cells were stimulated with 5 ng/ml TGF-β1 for 48 h. The cells were fixed for immunocytochemical analysis with the anti-α-SMA antibody and Alexa Fluor 546-conjugated second antibody. Nuclei were stained with DAPI. Scale bar, 50 μm. The results are presented as the mean ± S.E. ANOVA followed by Tukey's HSD test was performed for multiple comparisons. *, p < 0.05; **, p < 0.01.