microRNA (miR)-181c controls tumor necrosis factor-α production in primary rat microglial cells. (A-B) Primary rat microglia cells were transfected with miR-181c mimics or NC. After 48 hours, cells were harvested, and the TNF-α mRNA (A) and protein (B) expression levels were evaluated by real-time PCR and western blotting. (C-D) Primary rat microglial cells were transfected with miR-181c mimics or NC 24 hoursbefore activation by OGD. After 48 hours, the production of TNF-α was determined by ELISA (C). Cultured neurons were then grown in the microglia-conditioned medium (MCM) for 48 hours, and neuronal apoptosis was detected using Hoechst 33342 staining (D). (E-F) Primary rat microglia cells were transfected with miR-181c mimics or NC 24 hoursbefore activation by OGD. After 48 hours, the production of NO was determined by the Griess assay (E). The cells were harvested, and iNOS protein expression was evaluated by western blotting (F). The results are presented as the mean ± SE from three independent experiments. Ctrl represents the microglial cells that were not treated with oligonucleotide transfection; NC represents the negative control for the miR-181c mimics. *P < 0.05. NS, not significant.