Figure 2.

Enforced expression of MAZR partially restores impaired CTL differentiation in the absence of MAZR and Runx3. (A) Schematic figure showing the retroviral vector-mediated gene transduction of MAZR or Runx3 into WT and M/R3-cDKO^CD4 CTLs. See the Methods section for the detailed experimental procedure. (B) Histograms showing CD8α, Granzyme B, T-bet, Eomes and IFN-γ expression in GFP⁺ WT CTLs transduced with a control vector as well as in GFP⁺ M/R3-cDKO^CD4 CTLs transduced with control, MAZR- and Runx3-expressing vectors (6 days after in vitro anti-CD3/28 stimulation). T helper 2 (Th2) cells were included as negative controls for CD8α, Granzyme B, T-bet and Eomes stainings, whereas WT CTLs without restimulation were used as a negative control for IFN-γ staining. Vertical dotted lines indicate the peak of CD8α expression in WT CTLs or populations expressing the respective proteins based on the negative controls. (C) Diagrams showing the relative mean fluorescence intensity (MFI) of CD8α, Granzyme B, T-bet, Eomes and IFN-γ expression in GFP⁺ M/R3-cDKO^CD4 CTLs transduced with control, MAZR- and Runx3-expressing vectors. The MFI values of GFP⁺ WT CTLs transduced with control vector were set as 1, and are indicated as horizontal dotted lines. Each dot represents value from each experiment, and paired experiments are indicated with lines. A paired two-tailed Student’s t-test was performed for statistical analysis. The p-values were defined as following: *p < 0.05. Data are representative (B) or show the summary (C) of 3-4 independent experiments.