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. Author manuscript; available in PMC: 2018 Aug 16.
Published in final edited form as: Cell Rep. 2018 Jun 12;23(11):3262–3274. doi: 10.1016/j.celrep.2018.05.050

Figure 7. The Presence of Ezh2-Deficient Tregs Are Required to Promote Tumor Rejection.

Figure 7.

(A) Timeline for diphtheria toxin (DT) administration to FP3-DTR mice or tam administration to FP3-ER mice bearing MC38 tumors in (B), (C), and (D).

(B) Percentage of FOXP3+ Tregs of CD4+ T cells in lymph nodes.

(C) Representative pictures of mice temporally disrupted for EZH2 in Tregs (left) or depleted of Tregs (right).

(D) Absolute number of CD8+ T cells (top) and effector CD4+ T cells (eCD4+: CD4+Foxp3, bottom) in tumor, lung, and draining lymph node (dLN) of indicated mice.

(E) TRAMPC2 and MC38 tumor growth in FP3-DTR micewith orwithout DT treatment (time ofadministration indicated by shading). Inset numbers represent the number of mice that rejected tumors of total implanted.

(F) Scheme (top) and representative flow cytometry plot (bottom) illustrating chimeric female model fortransiently eliminating wild-typeTregs in the presence of Ezh2-deficient Tregs.

(G) MC38 tumor growth in indicated chimeric female mice treated with diphtheria toxin from tumor inoculation throughout.

(H) MC38 tumor growth in indicated chimeric female mice or FP3-DTR/FP3-DTR mice treated short term with diphtheria toxin from days—2 to 4 after tumor inoculation. Inset numbers represent the number of mice that rejected tumors of total inoculated in one of two experiments(see Figure S6 for cumulative results). p values indicate comparison between red (Ezh2fl/fl) and blue (FP3-DTR) lines, but comparison of red to black (Ezh2fl/+) lines at these three time points also was significant. No significant difference at any time point observed between black and blue lines.

Data represent means ± SEMs from two or more experiments; *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 from two-way ANOVA with Sidak’s multiple comparisons or by unpaired t tests. See also Figure S6.