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. 2018 Oct 3;115(42):E9859–E9868. doi: 10.1073/pnas.1805474115

Fig. 7.

Fig. 7.

RKIP competes with Gab2 to interact with the PI3K p85 subunit. (A) HEK293T cells were transfected with Myc-RKIP and Flag-Null, Flag-Lyn, Flag-PLCγ1, Flag-p85, or Flag-Akt plasmids; then they were immunoprecipitated (IP) with anti-Flag beads (M2 beads) and probed with anti-Myc or anti-Flag antibody. (Lower) Immunoblots analyzing whole cell lysates (WCL) without immunoprecipitation and (Upper) immunoblots analyzing immunoprecipitated flag-tagged proteins. (B) Flag-EGFP or Flag-p85 proteins purified from overexpressed HEK293T cells were mixed with recombinant GST-RKIP protein, and then the proteins were pulled down with M2 beads. (C) The lysates of the IgE−FcεRI-activated BMMCs were IP with anti-p85 or IgG (control) and probed with anti-RKIP or anti-p85 antibody. (D) Immunoblot of the HEK293T cells transfected with EGFP-Gab2 and Flag-p85 with or without Myc-RKIP, then were IP with M2 beads. (E) The lysates of IgE−FcεRI-activated RKIP WT or KO BMMCs were immunoprecipitated with anti-p85 or IgG (control) and probed with anti-Gab2 or anti-p85 antibody. Data are representative of three experiments.