Fig. 4

MiR-494 regulated Nrdp1˛levels in vitro and in vivo (a) Nrdp1 is a direct target of miR-494 in macrophage. The region of the Nrdp1 mRNA 3’UTR predicted to be targeted by miR-494 as indicated. b Luciferase activity assays using reporters with wild-type or mutant Nrdp1 3’UTRs were performed after cotransfection with miR-494 mimics or control in macrophage. c Macrophages were transfected with miR-494 mimics or inhibitors. After 24 h, cells were harvested, and miR-494 levels were evaluated by qRT-PCR. Transduction of miR-494 mimics enhanced miR-494 mRNA levels, while transduction of miR-494 inhibitors decreased miR-494 mRNA levels. d Macrophages were transfected with miR-494 mimics or inhibitors, and then were treated with PBS or erythrocyte lysates for 3 days. After then, neuron was treated with conditioned medium from treated macrophage. After 48 h, the Nrdp1 expression of macrophages was analyzed by western blot assays. e Intracerebroventricular injection of miR-494 mimics or inhibitors was administered 10 min after ICH. miR-494 mimics significantly attenuated Nrdp1 levels compared with control group. Experiments performed in triplicate showed consistent results. The differences were analyzed using ANOVA. *P < 0.05