Figure 7.

GLP1-mediated anti-apoptotic effects were abolished when Akt was inhibited. Cultured cardiomyocytes were incubated with PA (400 μM) for 24 h in the absence or presence of GLP1 (25 nM) alone, or in combination of GLP1 (25 nM) with an Akt inhibitor MK2206 (50 nM). (A) Apoptotic cardiomyocytes were examined by TUNEL staining (red, TUNEL and blue, DAPI; scale bar, 50 μm). (B) Numbers of apoptotic cells were quantified and expressed as the percentage of TUNEL-positive to DAPI-positive cells. (C) Levels of cleaved caspase-3 were analyzed by western blot and quantified by densitometry. Distribution of active b-catenin in cardiomyocytes was determined by immunostainning (red, b-catenin and blue, DAPI; scale bar, 10 μm) (D) and levels of cytosolic and nuclear b-catenin were further examined by western blot (E). Intensities were quantified and normalized against the level of GAPDH or histone-3 and expressed as fold changes of protein abundance under PA stimulus. Data are means±s.e.m. of three independent experiments. *P<0.05 and **P<0.01 vs PA; ^##P<0.01 vs PA+GLP1. PA, palmitate. A full colour version of this figure is available at http://dx.doi.org/10.1530/JME-15-0155.