Nuclear dye dapi

Cells were collected by cytospin on glass slides and fixed with 4% paraformaldehyde for 15 min. After washing in 10% PBS, the slides were permeabilized with 0.2% Triton X-100 in PBS and then were blocked with 4% bovine serum albumin (#A8020; Solarbio, Shanghai, China) for 1 h at room temperature. After incubated with LC3A/B antibody (#12741, 1:1000, Cell Signaling Technology, Danvers, USA) overnight at 4 °C, the slides were washed with PBS and then incubated with Alexa Fluor 647s antibody (#bs-0295G-AF647, Bioss, Woburn, USA). The cells were stained with nuclear dye DAPI (#C0080; Solarbio, Shanghai, China). All the images were captured with the fluorescence microscope, and representative images were shown.

Paraffin-embedded tissue sections were blocked with BSA for 2 h at room temperature and incubated with rabbit anti-LC3B (1:100; NB100-2220, NOVUS, Littleton, CO, USA) or PINK (1:100, AF7755, Beyotime, Shanghai, China) primary antibodies overnight at 4 °C, and then incubated with rabbit fluorescent secondary antibodies (1:200; Invitrogen, Carlsbad, CA, USA) for another 1 h at 37 °C. Cells were stained with the nuclear dye DAPI (1:1000, Solarbio, Beijing, China) for 5 min at room temperature and observed with a confocal microscope (Zeiss LSM 710 META, Jena, Germany).