Plasmid DNA Transfection and AAV Production

The plasmid DNA were obtained from ViGene Biosciences (Rockville, MD, USA). Endotoxin-free plasmid DNA was generated and purified by NucleoBond Xtra Midi Plus EF kit (Macherey-Nagel, Dueren, Germany). βTC cells were transfected with plasmid DNA containing Atg7 shRNA, or negative control using Lipofectamine 3000 (ThermoFisher, Waltham, MA, USA). The efficacy of knockdown was assessed. Adeno-associated virus (AAV) serotype 8 vectors were generated by polyethylenimine (PEI) transfection of HEK-293 cells as previously reported^{19 (link),20 (link),27 (link),28 (link)}. AAV was purified using discontinuous iodixanol gradients. Purified AAV vectors were filtered and stored at −80°.