Tg4<sup>KO</sup> mice show reduced induction of IL-10<sup>+</sup> T cells and tolerance-associated transcription factors following [4Y] treatment.

<p>(<b>A</b>) Representative flow cytometry plots and plots of all data (<b>B-G</b>) showing production of selected cytokines by splenic CD4<sup>+</sup> T cells from Tg4<sup>WT</sup> and Tg4<sup>KO</sup> mice treated with [4Y] or PBS following <i>ex vivo</i> restimulation with PMA and ionomycin. (<b>H</b>) Representative flow cytometry plots and plots of combined data (<b>I-J</b>) of FoxP3 and cMaf expression in splenic CD4<sup>+</sup> T cells from Tg4<sup>WT</sup> and Tg4<sup>KO</sup> mice treated with [4Y] or PBS. All flow cytometry data is gated on live (viability dye<sup>-</sup>) CD4<sup>+</sup> cells. (K-M) Expression of <i>Il10</i>, <i>cmaf</i> and <i>Nfil3</i> in magnetically-isolated CD4<sup>+</sup> splenocytes from Tg4<sup>WT</sup> and Tg4<sup>KO</sup> treated with [4Y], restimulated for 18 hours <i>ex vivo</i> with anti-CD3ε and CD28, as measured by RT-PCR. Plots F, G, K-M show data from one experiment with 3–4 mice per group. All other plots show the data from two experiments combined, with a total of 6–8 mice per group. All plots show the mean +/- SEM with each point representing data from one animal. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns p>0.05 assessed by ANOVA with Tukey’s correction for multiple comparisons (plots B-G and I-J). p values shown in K-M were calculated by two-tailed Student’s T test.</p>