(a) Naive CD4+ T cells from wild-type (Mir210+/+CD4-Cre), Mir210–/– (Mir210f/fCD4-Cre) or Hif1af/+Mir210f/fCD4-Cre mice were differentiated under TH17 skewing conditions (0.2 ng/ml of IL-6), followed by intracellular staining of IL-17A and IFN-γ. (b) Naive CD4+ T cells from wild-type (Mir210+/+CD4-Cre) or Mir210–/– (Mir210f/fCD4-Cre) mice were differentiated under TH17 skewing conditions with varying doses of IL-6 under normoxic or reoxygenetion conditions, followed by intracellular staining of IL-17A and IFN-γ. (c–d) Wild-type or Mir210–/– T cells were TH17 differentiated under reoxygenetion conditions as described in (a) for 3 d. Transcripts of miR-210 and Hif1a were detected by RT-PCR analysis; HIF-1α and GAPDH protein levels were detected by immunoblot analysis. (e–f) Quantitative RT-PCR analysis of HIF-1α target genes of the glycolytic pathway (Eno1, Glut1, Ldha, Pkm, Slc16a3 and Tpi) and TH17 signature genes (Il17a, Il17f, Rorc and Il23r) in wild-type or Mir210–/– CD4+ TH17 cells that were differentiated under reoxygenetion conditions. Relative expression is normalized to sno202 (miR-210) or β-2m (the rest of genes). Data are from one experiment representative of two (a right, d,e), three (c,f) or four (a left and middle, b) independent experiments. (mean and s.d. in b,c,e,f).