Fig. 3.

miR-212-5p was constantly downregulated after CCI and negatively regulated ferroptosis in neuronal cell lines. a Quantitative RT-PCR showing the relative expression levels of miR-212-5p in the cortex at 6 h, 12 h, 1d, 2d, 3d after injury (n = 9 for each group). b The HT-22 and Neuro-2a cells were treated with Rsl3 (3 μM) with or without a cell death inhibitor (ferrostatin-1, 1 μM; zVAD-fmk, 10 μM; necrosulfonamide, 0.5 μM) for 24 h. Data were analyzed using analysis of variance (ANOVA) with Tukey’s post hoc test. c Overexpression of miR-212-5p suppressed Rsl3-induced cell death in HT-22 and Neuro-2a cells. d Inhibition of endogenous miR-212-5p enhanced Rsl3-induced cell death in HT-22 and Neuro-2a cells. Indicated cells were treated with Rsl3 (3 μM) for 24 h. The transfection efficacy of overexpression (e) or inhibition (f) was confirmed by qRT-PCR analysis. Cell death was assayed using the Cytotoxicity Detection kit. Data shown represent mean ± SEM from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001. CCI, controlled cortical impact