Gabarapl2

After incubation cells were lysed directly into sample buffer (50 mm Tris-HCl buffer pH 6.8, 2% SDS, 10% glycerol, 0.005% bromophenol blue) containing 100 mm maleimide to preserve protein oxidation. Protein samples were resolved on SDS-PAGE gels and then transferred to polyvinylidene difluoride membranes by western blotting (TransBlot Turbo, Bio-Rad). Membranes were blocked in milk dissolved in phosphate-buffered saline (PBS) supplemented with 1% Tween before incubation with primary antibodies. Membranes were developed using enhanced chemiluminescence substrate (ThermoFisher) and detected using photographic film, which was developed using an automatic processor (Fuji RG II). Samples were immunoblotted for LC3 (1:1000, Cell Signaling), Atg3 (1:5000, Abcam), Atg7 (1:1000, Cell Signaling), Actin (1:10,000, Sigma), Atg5 (1:1000, Cell Signaling), Atg12 (1:1000, Cell Signaling), Atg4B (1:1000, Cell Signaling), Atg10 (1:1000, Abcam), GABARAPL1 (1:1000, Cell Signaling), GABARAPL2 (1:1000, Cell Signaling), cGMP-dependent protein kinase (PKG, 1:1000, ENZO Life Sciences), Myc-tag (1:1000, Cell Signaling), p62 (1:500, Cell Signaling), caspase-3 (1:1000, Abcam), peroxiredoxin-SO3 (1:500, AbFrontier) and GFP (1:2000, Miltenyi Biotec). All uncropped western blots can be found in Supplementary Fig. 6.