Fig. 9
IL-15Rα is differentially associated with the IL-15 receptor β/ γ upon engaging extracellular and intracellular IL-15. (A), CD3+ T cells isolated from PBMCs were stimulated with the IL15/IL15Rα complex (100 ng/mL) for 3 days. The expression of Ki67, GZMB, and perforin was assessed by flow cytometry. (B), A Transwell migration assay was performed on A549 and PC9 cells in the presence or absence of the IL15/IL15Rα complex (100 ng/mL). The data are presented as the means ± SDs. Scale bar, 100 μm. (C and D), Antibodies against IL15 (C) or IL15Rα (D) were added to the lysates of A549 and PC9 cells; (E), Antibodies against IL15 were added to the lysates of IL-15- overexpressing A549 and PC9 cells. The proteins pulled down by the antibodies were analyzed by immunoblotting. The protein levels of IL15Rα, IL2/IL-15Rβ, and IL2/IL-15Rγ were measured. (F), Transwell migration assays were performed on IL-15-overexpressing A549 and PC9 cells that were transfected with siRNA-IL15Rα or siRNA-Ctrl for 48 h. The data are presented as the means ± SDs and were analyzed via one-way ANOVA. Scale bar, 100 μm.**, p < 0.01; ***, P < 0.001; ****, p < 0.0001. (G), Antibodies against IL15Rα were added to the lysates of A549 cells treated with or without IL-15 for 5 or 10 min after which the proteins pulled down by the antibodies were analyzed by immunoblotting. The protein levels of IL15Rα, IL2/IL-15Rβ, and IL2/IL-15Rγ were measured