FIG 9.

Runx3 is required for GzmB expression but not IFN-γ production. CD45.1 and CD45.1/2 splenocytes were magnetically purified for CD4 T cells and electroporated to incorporate Cas9 and Runx3 sgRNA or control nontargeting sgRNA. Totals of 2 × 10⁶ to 4 × 10⁶ electroporated cells were mixed at a 1:1 ratio, adoptively transferred into CD45.2 recipient mice, and infected with ECTV. The spleens and livers were collected at 8 dpi for analysis. (A) Depiction of experimental design. (B) Representative flow plot of CD44 by GzmB expression by CD4 T cell populations. Samples were concatenated across each cell population group. (C) Percentage of GzmB⁺ for CD4 T cell populations. (D) Frequency activated CD44⁺ of CD4 T cell populations. (E) Percentage IFN-γ⁺ or IFN-γ⁺ TNF-α⁺ for CD4 T cells for each CD4 T cell population. Asterisks (*) represent statistical differences compared to the control sgRNA group, whereas number symbols (#) denote differences compared to the endogenous group. Compiled data are displayed as means ± the SEM from three independent experiments (n = 14).