PI3 kinase assays were performed after immunoprecipitation with antibodies against IRS2 (Cell Signaling L1326) and carried out as previously described (Wang and Summers, 2003). Immunoblotting for pAkt (Ser473, 4060) and total Akt (2920) (Cell Signaling Technology, Inc.) was performed as previously described (Kusminski et al., 2012).
