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

FIGURE 5.

FIGURE 5.

GLP-1 and forskolin but not high glucose nor insulin mimics the Akt kinase stimulatory effects of GIP. A, INS-1 cells were pretreated with DMSO (control) or 15 μm LY294002 for 30 min and then treated ± 10 nm GIP, 10 nm GLP-1, or 500 pm insulin (all with 3 mm glucose) or 16 mm glucose for 15 min, and Western analysis was performed on Akt KA assays with indicated antibodies. IP, immunoprecipitate. B, shown is the mean change ± S.E. in Akt activity (P-GST-GSK3/Akt) relative to DMSO (n = 4). #, p < 0.05 versus control without LY294002. C, INS-1 cells were pretreated with DMSO (control) or PI3K/Akt signaling inhibitors (15 μm LY294002 or 5 μm Akt VIII) for 1 h and then treated with ±1 μm forskolin for 15 min, and Western analysis was performed on cell lysates or Akt KA assays with indicated antibodies. D, shown is the mean change ± S.E. in Akt activity (P-GST-GSK3/Akt) relative to DMSO control (n = 4). #, p < 0.05 versus DMSO control. Anti-β-actin and anti-GST (GST-GSK3) blots were internal controls.