Figure 5. IL10 induces oncogenic signaling through inducing the interaction of IL10RA with phospho-Src.

A. Effect of IL10 knockout on EGFR^L858R and Kras4b^G12D-induced signaling. Lungs were homogenized for Western Blotting using the indicated antibody. Lower panel is the quantitative result. Data were expressed as mean±s.e.m. (*p<0.05, **p<0.01, ***p<0.001). B, C, D. After treatment with IL10 for 24 h, cell harvests were prepared for Western blotting with indicated antibodies. The lower panel of (C) and (D) is quantitative result. E. The immune-complex immunoprecipitated by the anti-Src antibody was analyzed by Western blotting using the indicated antibody. F. After transfection with wild-type HA-Src or kinase-dead HA-Src (aa 1-249), for 24h, H1299 cells with or without IL10 treatment were harvested for immunoprecipitation by the anti-HA antibody, and analyzed by Western blotting. Upper panel is the domain illustration of HA-Src. G. After transfection of pHA-Src and pHA-Src(1-249), and IL10 treatment, cellular extracts were prepared for Western blotting using indicated antibodies. Lower panel is the quantitative result for phospho-JAK1 and phospho-Akt. Data were expressed as mean±s.e.m. *p<0.05. H, I. After transfection with indicated plasmids and IL10 treatment, cellular extracts were prepared for immunoprecipitation using the anti-HA or anti-GFP antibody. The immunoprecipitated complex was analyzed by Western blotting.