Lentivirus was produced by transfection of CSII-CMV-MCS-IRES2-Venus, CSII-CMV-Cre-HA-IRES2-Venus, CSII-CMV-FLAG-GABARAPL2 ΔN-IRES2-mRFP, CSII-CMV-FLAG-GABARAPL2 ΔN-IRES2-Cre-HA, CSII-CMV-FLAG-p62 plasmids, or pSIH1-copGFP/mRFP plasmids with shRNA sequences (table S1), with pCAG-HIVgp and pCAG-VSV-G plasmids into Lenti-X 293 T cells grown on 10-cm dishes and maintained in 8 ml of DMEM supplemented with 10% FBS and 1% penicillin/streptomycin mix at 37°C. Cultured medium was collected 2 days after transfection and filtered through a 0.45-μm syringe filter, and then lentivirus was precipitated using Lenti-X Concentrator (631232, Clontech Laboratories Inc.) according to the manufacturer’s instructions. Precipitated lentivirus was resuspended in 250 μl of neuronal culture medium and kept at 4°C protected from light until use.

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