Figure 6

miR-124-3p directly targets ROCK1. (A) Oligonucleotides containing the predicted miR-124-3p binding sites in the 3′UTR of ROCK1 mRNA was synthetized, while mutations on the “seed” sequences are designed as below. Alignment between the predicted miR-34a target sites and miR-124-3p is marked with black color. (B) HEK293T cells were co-transfected with 50 nM of either miR-124-3p mimics or NC and 100 ng pmirGLO Dual-Luciferase miRNA Target Expression Vector with Wt or Mut 3′-UTR of ROCK1. The relative luciferase activity was measured 48 h after transfection. *P < 0.05. (C) The down-expression of ROCK1 was confirmed by western blotting, after miR-124-3p or siROCK1 transfected, GAPDH was used as control. (D) Quantitative real-time PCR analysis indicated that the relative mRNA level of ROCK1 was significantly decreased after miR-124-3p or RNAi treatment. GAPDH was used as control. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.