CD8+ iTregs contribute to the preserved GVL effect mediated by combinational therapy. BDF1 mice were transplanted as stated in Fig. 5. Fourteen days post HCT, spleen and liver mononuclear cells were isolated. Representative flow gating strategy (A) displays IFNγ secretion from Teffs and injected iTregs (H2Kd−). Percentages of indicated cell populations, IFNγ production (B), and absolute number are quantified (C). Cells were also stained for granzyme B and Fas-L, with a corresponding isotype control (D) representative histogram analysis of the isotype control, CD4+ singular or combination therapy. Graphical quantification of the mean fluorescence intensity for each molecule is displayed in (D), n = 4. *p < .05; **p < .01; ***p < .001. Error bars indicate the mean of standard error.