miR-132 targets Foxo3 and leads to increased IFN-γ, CCND1, and CCNE1 levels. A, Splenocytes obtained from mice were transfected either with miR-132 mimic (Mimic) or Mock transfection control (Mock) for 24 h. miR-132 and Foxo3 levels were determined by RT PCR and Western blot. B, Splenocytes were activated with (1 μg/ml) and for 24 h. Cells were then transfected with 100nM miR-132 inhibitor (Inhibitor) or Mock transfection control (Mock) for another 24 h. Foxo3 levels were determined via RT PCR and Western blot. For miRNA normalization, Snord96_a was used as internal control. For mRNA, β-actin was used as the internal control. C, Levels of Ifn-γ after transfection with miR-132 mimic and miR-132 inhibitor were assessed by RT PCR. D, Expression of cell cycle progression genes Ccnd1 and Ccne1 by RT PCR after transfection with miR-132 mimic and inhibitor. Data are represented as mean ± SEM from replicate samples. Statistical significance is indicated as *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.