Cells were seeded for microscopy on MatTek glass-bottom 35 mm dishes (MatTek, #P35G-1.5-14) and fixed with 4% formaldehyde in PBS for 10 min and then washed in PBS prior to imaging.
Freshly isolated whole hearts from α-MHC MitoTimer mice were fixed in 4% paraformaldehyde in PBS overnight, followed by an overnight incubation in 30% sucrose in PBS. The tissue was snap-frozen in OCT medium and 10 μm sections were stained with anti-LC3B/MAP1LC3B (1:100, Novus Biologicals #NB100-2220) primary and DyLight 350 conjugated anti-rabbit secondary antibody (Thermo Fisher–Pierce, #62270).
Fluorescence microscopy imaging was performed using a Keyence BZ-9000 microscope and analyzed with Keyence BZ-II Analyzer ver. 2.2. Confocal microscopy was performed using Leica TCS SP5 X White Light confocal microscope (Cedars Sinai Medical Center Microscopy Core). Images were collected and analyzed on Leica Application Suite X (LAS X) ver. 1.9.
Freshly isolated whole hearts from α-MHC MitoTimer mice were fixed in 4% paraformaldehyde in PBS overnight, followed by an overnight incubation in 30% sucrose in PBS. The tissue was snap-frozen in OCT medium and 10 μm sections were stained with anti-LC3B/MAP1LC3B (1:100, Novus Biologicals #NB100-2220) primary and DyLight 350 conjugated anti-rabbit secondary antibody (Thermo Fisher–Pierce, #62270).
Fluorescence microscopy imaging was performed using a Keyence BZ-9000 microscope and analyzed with Keyence BZ-II Analyzer ver. 2.2. Confocal microscopy was performed using Leica TCS SP5 X White Light confocal microscope (Cedars Sinai Medical Center Microscopy Core). Images were collected and analyzed on Leica Application Suite X (LAS X) ver. 1.9.