On day of transfection, transfect pCIG2-LAMP1-mNeon Green, pHaloTag-LC3B, and an autophagy regulating protein tagged with mcherry via lipofectamine 3,000 (ThermoFisher). Briefly, dilute plasmids in Opti-mem reduced serum medium and add P3000 reagent in accordance with manufacturer instructions. Also dilute lipofectamine 3000 reagent in optimem, then mix the dilute plasmid and lipofectamine solutions together. Incubate for ~10 minutes, then add dropwise to adhered cells.