Western blot analysis and immunoprecipitation were performed as previously described (18 (link)). For immunoprecipitations samples of 100–200 μg cell lysate protein in a 200–300 μl final volume of RIPA++ lysis buffer were incubated with appropriate antibodies (anti-HA cat # SC57592, lot #D1515 from Santa Cruz Biotechnology, anti-GFP cat #A11122, lot #1828014 from Invitrogen and anti-Myc Cat # 71D10 from Cell Signaling) overnight at 4 °C on an end-over-end mixer. Following 1 wash in RIPA++ buffer, they were used for lipid kinase activity assay or washed 3 more times prior to SDS PAGE.