Fig. 4.

IL-7 receptor expression and signaling and CDC42-PAK1 co-immunoprecipitation. A Surface expression of IL-7Rα (CD127) on CD4 + and CD8 + T cells from healthy controls (HC) and patients (P1, P2). B Flow-histograms depicting IL-7Rα expression on CD4 + and CD8 + T cells from patients and controls. In the upper panel HC and P1 follow the same expression pattern, in the lower panel P1 and P2 follow the same expression pattern. C,D Surface expression of IL-7Rα (CD127) in CD45RO + memory (C) and CD45RO- naïve subsets (D), from healthy controls (HC) and patients (P1 and P2). Only measurements > 1000 events for the IL-7Rα gate are depicted. E PBMCs stimulated with IL-7 for 0, 15 or 30 min were blotted for induction of phosphorylated STAT5, total STAT5 and GAPDH (loading control). F Quantification of E depicting ratio of pSTAT5/STAT5. G Co-immunoprecipitation of FLAG-CDC42 and PAK1 in HEK293T cells. CDC42 Lys16Glu, CDC42 WT and PAK1 were overexpressed in HEK293T cells as shown, and co-IP was performed by pulling down FLAG-CDC42 and performing WB for the immunoprecipitated PAK1. MFI: Median Fluorescence Intensity. Bars indicate median. In panel A, C, and D, 6 healthy controls were used, except for CD8 + CD45RO + IL-7R expression where 2 controls were uninformative due to insufficient gate events (< 1000 events/gate) Statistical comparisons: Mann–Whitney U test (2 groups), Kruskal–Wallis test (> 2 groups). *p < 0.05, **p < 0.01