Fig. 1

CAV3 was decreased in diabetic mice hearts and knock-down CAV3 induced mitochondria-dependent intrinsic apoptosis in cardiomyocytes. A Western blotting and associated quantitative analysis the CAV3 protein expression levels in the hearts of wt and db/db mice (n = 7 mice). B Representative immunohistochemical stains of CAV3 in the hearts of wt and db/db mice (n = 7 mice). C Representative Western blotting and quantitative analyses for CAV3, Bax, Cleaved caspase 3 and Bcl2 protein levels in cardiomyocytes treated with or without HGHF and with or without CAV3 downregulation (n = 4). D Representative Western blotting and quantitative analyses for the content of cytochrome C in cytoplasm treated with or without HGHF and with or without CAV3 downregulation (n = 3). E Flow cytometry and quantitative analysis of apoptosis cells by AnnexinV-FITC and propidium iodide (PI) staining in cardiomyocytes treated with or without HGHF and with or without CAV3 downregulation in vitro (n = 5). F Flow cytometry and quantification analysis of mitochondrial membrane potential by JC-1 in cardiomyocytes treated with or without HGHF and with or without CAV3 downregulation. High levels of green fluorescence (x-axis) represent reduced mitochondrial membrane potential (ΔΨm) and high levels of red fluorescence (y-axis) represents increased ΔΨm. A decrease in the red/green fluorescence is indicative of loss of ΔΨm (n = 6). G Normalized ATP production levels in cardiomyocytes treated with or without HGHF and with or without CAV3 downregulation in vitro (n = 6). H Representative confocal microscope of MitoSOX staining, immunofluorescence staining of 8-oxoG andquantitative analyses in cardiomyocytes treated with or without HGHF and with or without CAV3 downregulation (MitoSOX fluorescence, red; 8-oxoG fluorescence) (n = 5). I Immunofluorescence images of 8-oxoG and TOM20 in the AC16 cells treated with HGHF. AC16 cells were stained with 8-oxoG antibodies (Alexa Fluor 594), DAPI, and TOM20 (Alexa Fluor 488). Data are depicted as mean ± SD. *p < 0.05, **p < 0.01