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. Author manuscript; available in PMC: 2022 Mar 9.
Published in final edited form as: Immunity. 2021 Mar 2;54(3):514–525.e6. doi: 10.1016/j.immuni.2021.02.015

Figure 5. miR-221 and miR-222 target Maf and Il23r to regulated Th17 response.

Figure 5.

(A) Overlap of curated Th17-signature genes (Ciofani et al., 2012) and predicted target genes for miR-221 and miR-222 (microRNA.org: (Betel et al., 2008)). (B) HEK293 cells were transfected with luciferase reporter fused to 3’ UTR of Il23r or Maf together with miR-221 plus miR-222 or control miRNA (random sequences). Luciferase activity was evaluated at 24 hr post-transfection. Data are representative of 2 independent experiments. (C) Flow cytometric analysis of c-Maf in WT and miR-221/222-KO Th cells, gated according to Figure 3A. Data show representative flow cytometry plots and the pooled data with statistical evaluation. (D-E) Ex vivo intestinal lymphocytes were stimulated in vitro with IL-23 or IL-23 and IL-1β for 6 hrs and subjected to flow cytometric analysis to measure the induction of c-Maf expression in Th17 cells (D; 3-4 mice/group) and the frequency of IL-17-producing RORγt+ CD4+ T cells in WT and miR-221/222-KO mice (E; 6-7 mice/group). See also Figure S5.