Fig. 5

miR-185 regulates ITGB5 expression. a Potential miRNAs of ITGB5 as predicted by TargetScan and miRanda. b The 3′UTR of ITGB5 was constructed into a pSICHECK2 vector and was cotransfected with miR-122, miR-486-5P, miR-139,miR-271 and miR-185 individually. The luciferase activities were measured. Data represent the mean ± SD of three independent experiments. *p < 0.05 and **p < 0.01 vs. control. c–d miR-185 and miR-271 were introduced into Huh-7 cells. The protein levels of ITGB5 were examined by western blotting. e–f miR-185 inhibitor was transfected into Huh-7 and MHCC-97 L cells. The protein levels of ITGB5 were examined by western blotting. g Schematic illustration of pSICHECK2-based reporter constructs used in luciferase assays to examine the 3′UTR activity of ITGB5. The different truncations were named F1, F2 and F3 as indicated. h The different truncations were respectively transfected into MHCC-97 L cells with or without miR-185 overexpression. The luciferase activity was measured. **p < 0.01 and ***p < 0.001 vs. control. i Sequences of ITGB5 3′UTR and the potential miR-185 binding site at the 3′ UTR of ITGB5 as well as nucleotides mutated in the 3′ UTR of ITGB5, where the red indicates the mutated region. j–k The wild type 3′UTR (WT) and mutated 3′UTR (MUT) of ITGB5 were transfected into Huh-7 and MHCC-97 L cells with or without miR-185 overexpression. The luciferase activities were measured.**p < 0.01 and ***p < 0.001 vs. control. l The wild type 3′UTR (WT) of ITGB5 was transfected into Huh-7 and MHCC-97 L cells with or without miR-185 inhibition. The luciferase activities were measured.**p < 0.01 vs. control