Analysis of autophagy flux in ATG5 KO cells. (A) ATG5 KO efficiency was examined by WB. (B–D) HEK293A reporter cells were transduced with viruses carrying sgRNA targeting ATG5. These cells were then treated with amino acid–free media for 3 h (B), tunicamycin (10 µg/ml) for 6 h (C), or clofibrate (1 mM) for 6 h (D). Next, they were examined using FACS. Histogram overlays compare either GFP or DsRed signals of the treated ATG5 KO reporter cells with those signals of the untreated ATG5 KO ones. Source data are available for this figure: SourceData FS1.