FIG 7.

CD69 downstream signaling regulates miR-155, STAT5, and socs-1 expression in Treg cells. (A, left) Representative plots of CD69 expression in sorted mouse Foxp3-mRFP/cd69^+/+ tTreg cells treated with anti-CD69 Ab 2.2 or the 2.8 isotype control antibody. (Right) CD69 expression after Ab treatment determined by FACS analysis. Bars correspond to the means ± standard deviations of data from one representative experiment of four. (B) qPCR analysis of mmu-miR-155 expression in sorted CD69⁺ or CD69⁻ Foxp3-mRFP⁺ tTreg cells after Ab treatment. Results are normalized by sno135 snRNA expression, and expression is relative to that in 2.8-treated CD69⁺ cells. (C, left) Representative histogram showing the levels of STAT5 phosphorylation in iTreg cells from cd69^+/+ or cd69^−/− reporter mice treated with anti-CD69 Ab 2.2 or the 2.8 isotype control Ab. (Right) Quantification of STAT5 phosphorylation levels shown as geometric mean fluorescence intensities. (D) socs-1 transcriptional levels analyzed by qPCR. Data shown in panels A and B are derived from 3 independent sortings/experiments (3 animals per sorting), and data shown in panels C and D are iTreg cells differentiated from at least 4 mice per group. Data were analyzed by 1-way ANOVA and Bonferroni's posttest (B). CD69 expression after Ab treatment was analyzed by a t test (A). *, P < 0.05; **, P < 0.01; ***, P < 0.001 (Student's t test).