Fig. 4

Correlations between miR-124 and Bim were explored. (A) HBMECs were transfected with miR-124 mimic, and the increased miR-124 level was confirmed by qRT-PCR assay. NC, negative control. *P < 0.05. (B) HBMECs were transfected with miR-124 mimic, and the inhibited Bim mRNA level was confirmed by qRT-PCR assay. NC, negative control. *P < 0.05. (C) The impact of miR-124 on Bim was identified by western blot assay. Compared with control group, the overexpression of miR-124 down-regulated Bim protein level. NC, negative control. (D) The predicted 3′-UTR of Bim contains two miR-124 seed sequence (465-471 and 2394-2400) and deletion of the putative miR-124 binding seed sequence. (E) MiR-124 target gene was validated by flow cytometry analysis. A significant reduction in luminescence was displayed in HEK 293T cells transfected by miR-124 mimic and the Bim 3′-UTR wild-type reporters, whereas no significant difference was obtained in the mutant version of both sites (Bim-mut). Furthermore, HEK 293T cells transfected with miR-124 mimic and Bim-mut 1 or Bim-mut 2 exhibited an evident shrinkage. ns, no significance. *P < 0.05