Figure 5.

Altered in vitro CTL differentiation by loss of MAZR and/or Runx3 is largely due to CD8⁺ T cell lineage-intrinsic defects. (A) Histograms showing the expression of indicated proteins in the mutant CTLs (6 days after in vitro anti-CD3/28 stimulation). T helper 2 (Th2) cells as well as WT CTLs without restimulation were included as negative controls for the stainings of Granzyme B, T-bet and Eomes as well as IFN-γ, respectively. Vertical dotted lines indicate the peak of CD8α expression in WT CTLs or populations expressing the respective proteins based on the negative controls. Due to the appearance of Runx3-expressing cells within Runx3-cKO^E8I and M/R3-cDKO^E8I CTLs (that had escaped from Cre-mediated excision of the Runx3 allele), the expression pattern in the two mutant cells were analyzed in the Runx3-negative population by including intracellular staining of Runx3 protein. (B) Diagrams showing the relative mean fluorescence intensity (MFI) of indicated proteins in the mutant CTLs (the values of WT cells set as 1). For the comparison of values between three mutant groups, a one-way ANOVA analysis was performed. The p-values were defined as following: *p < 0.05; **p < 0.01. For the comparison of the relative MFI values between WT (set as 1) and individual mutant CTLs, a one-sample t-test was performed. The p-values were indicated above the diagrams. Data are representative (A) or show the summary (B) of 4-6 mice analyzed in 4-6 independent experiments.