Fig. 4

TGFBR1 and SMAD2 are the targets of miR-27b. a The binding sites of miR-27b on the 3’-UTRs of TGFBR1 and SMAD2 as predicted by TargetScan. b 3’-UTR luciferase reporter assay. HEK 293 T cells were transfected with TGFBR1-UTR or SMAD2-UTR luciferase vector together with the miR-27b expression plasmid or vector control (CON). pmiRGLO is the empty vector without any 3’-UTR sequences. Luciferase activities were determined 48 h post-transfection. The fold changes were relative to CON for each reporter construct. Data shown are means ± S.E *P < 0.05, **P < 0.01 n = 4. c, d TGFBR1 and SMAD2 protein levels in miR-27b overexpressing fibroblasts. LL29 fibroblasts were infected with a miR-27b lentivirus or virus control (VC) at a MOI of 50. After 72 h, the cells were lysed for Western blotting. Protein levels were quantitated by Image J software and normalized to β-actin. The fold changes were relative to VC. Date were represented as means ± S.E., *P < 0.05, n = 3–4. ANOVA, followed by Turkey’s test