Extended Data Fig. 8: ATG2 overexpression induces toxicity through its lipid transfer function.

a. Representative GFP immunofluorescence images and western blots in ATG2A/B KO HeLa transfected with indicated constructs showing that the localization and expression of GFP-ATG2-LTD is similar with GFP-ATG2A-WT. Scale bar: 10 µm. This experiment was done twice. b. Representative brightfield and fluorescence images of the phenotypic abnormalities found in clutches injected either with 200 pg GFP empty vector or GFP-ATG2A WT constructs compared to their uninjected siblings at 48 h.p.f. Scale bars: 2 mm for clutches, 1 mm for individual embryos. N = 4 independent experiments. c. Primary neurons were pretreated with the following drugs: DMSO (control), 10 µM Z-VAD-fmk, 20 µM necrostatin, 500 nM liproxstatin-1 (Lip-1), 5 µM ferrostatin-1 (Fer-1), 10 µM Mitotempo (MT), 10 nM ubiquinol (QH2), 5 µM PISD inhibitor (iPISD) for 12 hours before 24-hour scramble and smartpool shWIPI4 lentivirus treatment together with the above drugs. Cell cytotoxicity (n = 3 biologically independent experiments) was measured by LDH release assay. Data are presented as normalized mean ± SD. ns., not significant, *P < 0.05, **P < 0.005. Two-tailed one-sample t-test when compared with scr DMSO and two-tailed paired t-test with shWIPI4 DMSO. d. The knockdown efficiency of WIPI4 (n = 3 independent experiments) in Fig. 6j and Extended Data Fig. 8c. Source numerical data and unprocessed blots are provided.