Fig. 4: Mechanism of the enhanced survivability of γδT cells under IL-15 culture condition.

When γδT cells were cultured for 14 days, cytokines were withdrawn for 72 h and γδT cells were collected and analyzed. IL-2-fostered γδT cells as a control culture condition. A Flowchart of the experimental setup to study potential the mechanism of IL-15 to enhance the survivability of γδT cells. B Western blotting was performed to analyze the expression of Mcl-1 and Bcl-2 (HC, n = 8; NB, n = 9). Data were mean ± SEM from five independent experiments. Statistical analysis was performed using one-way ANOVA and Dunnett’s multiple comparisons. ***, p < 0.001. C, D γδT cells were treated with AT-7519 for 24 h and representative results were shown for the Western blotting and the flow-cytometric analysis. E Annexin V⁺ γδT cells were analyzed when γδT cells were treated with AT-7519 for 24 h (HC, n = 11; NB, n = 5). Data were mean ± SEM from three independent experiments. Statistical analysis was performed using one-way ANOVA and Dunnett’s multiple comparisons. *, p < 0.05; ***, p < 0.001. F Western blotting was performed to analyze the expression of STAT5 and ERK and the phosphorylation levels of STAT5 and ERK. The protein band intensity was quantified and analyzed by ImageJ (HC, n = 8; NB, n = 9). Data were mean ± SEM from four independent experiments. Statistical analysis was performed using one-way ANOVA and Dunnett’s multiple comparisons. *, p < 0.05; ***, p < 0.001.