Interleukin-4 Regulates Eomesodermin in CD8+ T Cell Development and Differentiation
Figure 3
Akt is required for IL-4 regulation of Eomes in CD8SP thymocytes.
A) Top, Flow cytometric analysis of intracellular Eomes expression in CD8SP thymocytes from d8 SLP-76 Y145F fetal thymi after culture in the absence or presence of AKTi-1/2 (5 µM) for the last 48 h. Bottom, Percentage of Eomes+ CD8SP among total CD8SP thymocytes in the indicated populations (n = 9, 2 independent experiments). B) Top, Flow cytometric analysis of intracellular Eomes expression in WT TCRβ+ CD8SP thymocytes after culture in the indicated conditions. Bottom left, Relative Eomes expression from cDNA of sorted CD8SP WT thymocytes after cultured in the indicated conditions, set relative to matched cells cultured in media alone. Bottom right, Percentage of Eomes+ cells among total CD8SP thymocytes (n = 5, 2 independent experiments). C) Top, Flow cytometric analysis of IL4Ra on CD8SP cells from WT thymocytes cultured as above. Bottom, percentage of IL4Ra+ cells among total CD8SP thymocytes in indicated conditions (n = 5/condition, 2 independent experiments). D) Flow cytometric analysis of surface CD44 on CD8SP thymocytes treated under indicated conditions as above. Right, percentage of CD44+ cells among total CD8SP thymocytes (n = 5/condition, 2 independent experiments). E) Top, Flow cytometric analysis of intracellular Eomes expression in TCRβ+ CD8SP thymocytes from WT fetal thymi after culture for 8d in the indicated conditions. Bottom, Percentage of Eomes+ cells among total CD8SP population (n = 8–9 per group, 2 independent experiments). Numbers in flow plots represent the percent of the gated population. Graphs show the average percentage of the indicated population and standard error of mean. Statistical significance calculated using Student’s t-test.