Research Paper Volume 12, Issue 11 pp 10441—10456

Figure 7. ROCK1 was a direct target of miR-34-5p and mediated the anti-fibrotic function of miR-34-5p. (A) The predicted binding sites of ROCK1 and miR-34-5p. (B) Luciferase reporter activities of chimeric vectors carrying the luciferase gene and a fragment of the 3’ UTR of ROCK1 containing the wild type or mutant miR-34-5p binding sites. Data was presented as mean ± SEM; two-tailed t test was used for the statistical analysis. n=5 independent cell cultures. (C, D) qRT-PCR analysis showing that overexpression of miR-34-5p inhibited the mRNA level of ROCK1 and AMO-34-5p transfection elevated the mRNA level of ROCK1; GAPDH mRNA served as an internal control. Data was presented as mean ± SEM; one-way ANOVA was used for the statistical analysis. n=5 independent cell cultures. (E) Protein levels of collagen 1 and α-SMA were measured by western blotting; GAPDH served as an internal control. Data was presented as mean ± SEM; one-way ANOVA was used for the statistical analysis. n=6 independent cell cultures. (F) Representative images of immunofluorescence staining showing that overexpression of miR-34-5p diminished fibroblast-myofibroblast transition induced by forced expression of ROCK1. Scale bars represented 50 μm. **P<0.05.