Figure 4

PI3Kα and γ isoforms associate with CNGA1 through their RA domain. The physical interaction of these domains was analyzed by reciprocal co-immunoprecipitations. The Flag-tagged RLD-CNGA1 domain and Myc-tagged PI3Kα and γ RA domains were transiently expressed in HEK-293 T cells. Expressed proteins were subjected to immunoprecipitation with either anti-Flag or anti-Myc antibodies or IgG. Anti-Flag and normal IgG IPs were subjected to immunoblot analysis with anti-Flag (A) and anti-Myc (B) antibodies along with Flag- and Myc-expressed protein lysates, whereas anti-Myc and normal IgG IPs were subjected to immunoblot analysis with anti-Myc (D) and anti-Flag (E) antibodies along with Myc- and Flag-expressed protein lysates. The relative binding was expressed as percentage of the RA-PI3K and RLD-CNGA1 and vice versa in each case (C, F). Data are mean ± SD, n = 3, *p < 0.001.