2.3　Immunofluorescence staining

Cells were seeded in 12-well culture plates on the cover glass，and treated as previously reported［17］.Briefly，cells were fixed with 4% paraformaldehyde，then washed with PBS twice.After being blocked with 10% FBSin PBS for 20min，cells were incubated with antiLC3 primary antibody for 1 h and secondary antibody Alexa Fluor546 goat anti-rabbit(Invitrogen，Carlsbad，USA)for 1 h，then washed with PBS three times.Imaging was performed with a 60×oil objective lens by Olympus FV1000 confocal microscope(Olympus，Tokyo，Japan).Each experiment was repeated three times.(https://www.daowen.com)