Fig. S3.

IFN-γ treatment increases LC3 puncta formation in HC04 cells. (A–C) HC04 cells were transfected with cDNAs encoding RFP-GFP-LC3. At 48 h posttransfection, they were treated with IFN-γ for 4 h in the presence or absence of 3-MA. RFP⁺GFP⁺-LC3 (autophagosomes) and RFP⁺GFP⁻-LC3 (autolysosomes) were quantified by determining the percentage of puncta-containing cells among at least 100 cells per condition from three independent experiments. Only puncta ≥0.25 µm in size were counted. The number of puncta per cell was also examined in Z-stack images of at least 30 cells per condition per independent experiment. Data are mean ± SEM. The results are expressed relative to the full control. (Scale bar: 5 µm.)