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. 2009 Apr 17;284(16):10764–10773. doi: 10.1074/jbc.M809116200

FIGURE 3.

FIGURE 3.

Phosphorylation of Akt at threonine 308 or serine 473 are not required for the activation of Akt by GIP. A, illustration of HA-Akt1 constructs and the respective point mutations used for this study. PH, pleckstrin homology domain; HM, hydrophobic motif. B, Western blot showing representative levels of HA-Akt1T308A/S473A protein in transfected cells relative to endogenous Akt in untransfected cells, as well as the purity of Akt1T308A/S473A immunoprecipitates. C, INS-1 cells transfected with HA-Akt1308/473 or HA-Akt1179 and 36 h later treated with ±10 nm GIP or 10 nm IGF-I for 15 min. Western analysis was performed on Akt KA assays with indicated antibodies. D and E, shown is the mean change ± S.E. in Akt activity (P-GST-GSK3/Akt) relative to Basal (n = 4) for INS-1 cells transfected with HA-Akt1308/473 (D) or HA-Akt1179 (E). #, p < 0.05 versus Basal. Anti-GST (GST-GSK3) blots were internal controls.