Figure 4.

miR-22 suppresses IL17 signaling in the T cells by reducing the recruitment of HIF1α/RORγT/STAT3 in the Il17a promoter

(A) The fold changes of RA signaling and Th17/Treg-related genes in hepatic T cells were quantified by RT-PCR and are shown in the heatmap. Hepatic T cells were isolated from livers of healthy, HCC, and miR-22-treated HCC mice followed by flow cytometry (n = 3). (B) Representative flow cytometry plots and percentage of Th17 (CD4⁺IL17A⁺), Tc17 (CD8⁺IL17A⁺), Th1 (CD4⁺IFNγ⁺), and Treg (CD4⁺CD25⁺FOXP3⁺) T cells in studied groups (n = 6). (C) ChIP-qPCR using anti-HIF1α, anti-RORγt, and anti-STAT3 antibodies in hepatic T cells. Hepatic T cells that were isolated from three mice for each studied group were subjected to ChIP assay. The primers for amplifying non-binding regions were used as a negative control. The regulatory regions of the Il17a and Rorc genes are shown with the binding locations of the indicated proteins. The numbers are relative to the transcription start site. Binding enrichment was expressed relative to the IgG-negative control. CNS, conserved non-coding sequence. Data represent mean ± SD, ∗∗p < 0.01; ∗∗∗p < 0.001 by one-way ANOVA (B and C).