Figure 11. Ptpn2 haploinsufficiency promotes conversion of RORγt⁺ effector Tregs.

(A) Kinetics of IL-17A⁺ exTreg generation during in vitro stimulation of sorted Ptpn2^+/+ FoxP3^eGFP SKG Tregs (n = 3) with IL-6 (50 ng/ml) and anti-CD3/CD28–coated beads. (B and C) In vitro conversion of Ptpn2^+/+ (n = 6) and Ptpn2^+/– (n = 5) effector and resting SKG Tregs into IL-17⁺ exTregs. (D) Generation of FoxP3^–RORγt⁺ exTregs (gated as in A) from Ptpn2^+/+ and Ptpn2^+/– effector and resting SKG Tregs. (E and F) Inhibition of JAK1/2 signaling using ruxolitinib during in vitro conversion of resting SKG Tregs. (E) Gating strategy for evaluation of RORγt⁺ expressing Ptpn2^+/+ (n = 7) and Ptpn2^+/– (n = 6) cells after 72 hours of culture. (F) Effect of JAK1/2 inhibition on upregulation of RORγt⁺ on live cells (top left, dot plot), generation of IL-17⁺FoxP3^– exTregs (top right, dot plot and histograms), and loss of FoxP3 within the RORγt⁺ population (bottom dot plot and histograms). Compiled data from 2 experiments are presented (C–F). Each symbol represents an individual mouse. Bars represent mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 by unpaired t test (C and D) or 2-way ANOVA (F).