Extended Data Fig. 5: Autophagy initiation hubs are conserved in human cells.

a, GFP–Atg11 Vac8-mCherry cells expressing endogenous BFP–Ape1 and copper-inducible untagged Ape1 were grown to mid-log phase in the presence of 50 µM CuSO₄. The fluorescence profile of GFP (green) and mCherry (magenta) was measured along the Ape1 condensate surface. Images of one out of three independent experiments are shown. Scale bar: 2 µm. b, Cells expressing Ede1–BFP, mScarlet–Atg11, and Vac8-mNeon were grown to mid-log phase. The fluorescence profile of mNeon (green) and mScarlet (magenta) was measured along the Ede1 condensate surface. Images of one out of three independent experiments are shown. Scale bar: 2 µm. c, GFP–Atg11 or GFP–Atg11 vac8∆ cells expressing endogenous BFP–Ape1 and copper-inducible untagged Ape1 were grown to mid-log phase in the presence of 50 µM CuSO₄. The fluorescence profile of GFP (green) was measured along the Ape1 condensate surface. Images of one out of three independent experiments are shown. Scale bar: 2 µm. d, GFP–Atg11 cells expressing endogenous BFP–Ape1 and copper-inducible untagged Ape1 were grown to mid-log phase in the presence of 50 µM CuSO₄. Atg11 rearrangement was monitored after rapamycin treatment. Quantification: BFP–Ape1 structures with rearranged Atg11 clusters. Scale bar: 2 µm. Data are mean values (n = 50 structures per condition and replicate, three biological replicates). Circles: mean values of each replicate, bars: mean. Statistical analysis: two-way ANOVA followed by Sidak’s multiple comparison test. P values: rich P < 0.0001, rapa P = 0.0005, rich(1-2):rapa(1-2) P < 0.0001, rich(>2):rapa(>2) P < 0.0001. e, Overlays of maximum intensity projection of a GFP–Atg19 cryo-fluorescence stack on 470x (left) and 6500x (right) magnification TEM images used to select acquisition region for tomogram shown in Fig. 4e. Scale bars: 10 µm (left) and 2 µm (right). One out of three identified examples is shown. Source numerical data are available in source data, arbitrary units (a.u.).