Figure 5.

GIMAP6 knockdown accelerates the PMA/ionomycin induction of T cell activation. A, Jurkat-derived cell lines (Ctl, KD1, and KD2) were treated with either PMA/ionomycin or an equal concentration of DMSO to determine the effect of GIMAP6 on T cell activation. To monitor the rate of cell apoptosis/death and T cell activation, PMA/ionomycin-treated cells were harvested at 3, 8, and 24 h and stained for the phosphatidylserine exposure marker Annexin V-FITC and for the T cell activation surface marker CD69-PE. The results were analyzed and quantified by FlowJo 7.6.1. B, the effect of GIMAP6 expression on PMA/ionomycin-induced cell death. All AnV+ cells were taken as apoptotic/dead cells, and the remaining AnV− cells were taken as live cells. C, the effect of GIMAP6 expression on PMA/ionomycin-induced T cell activation. To discriminate activated T cells from inactivated T cells across all live cells, the surface marker CD69 was used to calculate the percentage of CD69+/AnV−. D, the effect of GIMAP6 expression on PMA/ionomycin-induced IL-2 secretion. The data represent the average of at least two independent experiments with triplicate measurements (mean ± S.D.). P/I indicates that the cells were treated with both PMA (10 ng/ml) and ionomycin (1 μg/ml).