Figure 1: EphA2 associates with RCP.

(a) A2780 cells expressing GFP-RCP or GFP control were lysed in a buffer containing 0.15% Tween-20. GFP was immunoprecipitated from lysates using magnetic beads conjugated to an antibody recognizing GFP. Immunoprecipitated proteins were separated by SDS-PAGE. and visualized using Coomassie brilliant blue (CBB). GFP-RCP and α5-integrin were detected by immunoblotting (IB) as indicated. (b) A2780 cells were transfected with GFP and GFP-RCP or EphA2-GFP (upper panels) or were left untransfected (lower panel). Cells were lysed in a buffer containing 0.15% Tween-20 and lysates were incubated with magnetic beads conjugated to antibodies recognizing GFP (upper panels) or EphA2 (lower panel). The immunoprecipitates were analysed by immunoblotting using antibodies recognizing RCP and EphA2. (c,d) EphA2 was immunoprecipitated from H1299 (c) and PC3 (d) cells as for (b) and the immunoprecipitates analysed by immunoblotting for RCP, Rab11 and EphA2. Uncropped blots corresponding to the experiments presented in c,d are displayed in Supplementary Fig. 8.