Fig. 3
From: Dual roles of myocardial mitochondrial AKT on diabetic cardiomyopathy and whole body metabolism
Mitochondrial AKT1 promoted ATP synthase complex integrity. A Interruption of the formation of large ATP synthase complexes in TAM-CAMDAKT cardiac mitochondria. A sucrose gradient was used to fractionate mitochondrial protein complexes by size. Insulin stimulation in the TAM-CAMDAKT mice could not promote larger complex assembly as seen in the control mice. Representative blots are shown. Subunits b1 and c, required for ATP synthase anchoring of the cristae, were not associated with the larger complex in the TAM-CAMDAKT heart (n = 4 each group). B Interaction between ATP synthase complex and AKT1. ATP synthase subunits (ATP5A) and AKT1 from isolated mitochondria were co-immunoprecipitated. Representative blots are shown. ATP synthase subunit c and b1 did not associate with the AKT1 complex, which contains ATP synthase subunit alpha and beta, in TAM-CAMDAKT mitochondria but were evident in control mitochondria. (n = 3 each group). C Left, representative TEM images of CAMDAKT myocardial sections, 7 days after TAM or CO injection, showed loss of cristae in TAM-CAMDAKT mice compared to CO-CAMDAKT control animals (n = 2 each group). Scale bar = 1 μm (upper), 0.1 μm (lower). Right, quantification of TEM images. Each data point was the calculated average measurements of one image (CO:12 images, TAM:15 images). Cross-sectional areas of mitochondria, length of sarcomeres, M line widths, and intensity of myofibrils, a measure of myofibril density, were quantified. (All p < 0.0001)