Fig. 6.

WES analysis of Akt2 and Nck1. Capillary Western immunoassay (WES) was used to validate some of the proteins (Akt2 and Nck1) identified as differentially recruited by MS. Two hundred microliters of raft fraction collected from the OptiPrep gradient were precipitated with 10% TCA (final concentration) and analyzed by WES. Two biological replicates for T0 and T15 were used for Nck1 detection and one biological replicate for T0 and T15 was used to detect Akt2. Nck1 and Akt2 are detected in the condition T15 only. We used a pool T0 and T15 (rabbit IgG control) and a Master Mix (primary antibodies and secondary antibodies with no protein) to rule out nonspecific binding of antibodies. No bands were detected in these lanes.