Fig. 4

CAV3 interacted with mitochondrial complex I by anchoring NDUFA10 and maintaining its stability. A IP assay was carried out using CAV3 antibody and Coomassie Brilliant blue was stained to indicate CAV3 bands. B Through bioinformatics methods, NDUFA10 and TARDBP were overlapped from the LC–MS/MS analysis, DCM-related Genes (DisGeNet & Gencards) and Mitochondrial dysfunction-related genes (GeneCards). C, D IP assay was carried out using CAV3 or NDUFA10 antibody in cardiomyocytes. E Representative Western blotting assessing the purity of the mitochondria isolated. F–G IP assay was carried out using CAV3 or NDUFA10 antibody in mitochondria isolated from AC16 cells. H Immunofluorescence images of cardiomyocytes were stained with mitochondria-targeting dye (Mito-Tracker, Deep Red, Alexa Fluor 647), DAPI, NDUFA10 antibodies (Alexa Fluor 594), and CAV3 antibodies (Alexa Fluor 488). I Western blotting and quantitative analyses protein levels in cardiomyocytes of wt mice and db/db mice with or without CAV3 overexpression (n = 7 mice). J The levels of NDUFA10 mRNA in cardiomyocytes treated with or without HGHF and with or without CAV3 overexpression (n = 6). K CAV3 plasmid (pcDNA3.1-CAV3) was transfected into cardiomyocytes using Lipofectamine 2000 and treated with HGHF. The transfected cells were treated with 100 µmol/L cycloheximide for 0, 2, 4, 6, 8, 10 h. The protein levels of NDUFA10 were shown by a line chart (n = 3). L Cardiomyocytes were treated with or without HGHF, CHX (100 µmol/L), and MG132 (10 µmol/L) or CQ (50 µmol/L) for 24 h. Quantitative analyses of NDUFA10 protein levels among each group (n = 3). M Immunofluorescence images of NDUFA10 and lysosomes-targeting dye (Lamp1) in the cardiomyocytes treated with or without CAV3 overexpression. Cardiomyocytes were stained with NDUFA10 antibodies (Alexa Fluor 594), DAPI, and Lamp1 (Alexa Fluor 488). Data are depicted as mean ± SD. *p < 0.05, **p < 0.01