Figure 7: P733L mutation impairs JAK1 function.
From: Biallelic JAK1 mutations in immunodeficient patient with mycobacterial infection
(a,b) JAK1-deficient fibrosarcoma cells, U4A, were transfected for 24 h with vectors expressing sfGFP-fused JAK1WT, JAK1P733L, JAK1P832S or JAK1P733L/P832S. Then, cells were exposed to the indicated concentrations of IFN-α (a) or IFN-γ (b) for 15 min. Cells transfected with empty vector (e.v.) and parental, JAK1-competent, fibrosarcoma cells 2fTGH, served as negative and positive controls, respectively. *shows non-specific protein species detected by the antibody. Representative of four independent experiments. Fold change of band densitometry is indicated (numbers below bands and bar graphs in Supplementary Fig. 9).(c) Patient fibroblasts were transduced with vectors expressing JAK1WT, JAK1P733L, JAK1P832S or JAK1P733L/P832S and eGFP. Cells were stimulated with either IFN-α (100 ng ml−1) or IFN-γ (1 ng ml−1) or medium (−). One experiment. Fold change of band densitometry is indicated (numbers below bands).
