Extended Data Fig. 7: Mechanistic analysis of liposome uptake into droplets.
From: Phase separation promotes Atg8 lipidation and vesicle condensation for autophagy progression
a, Observation of BODIPY or NBD-PE-containing liposomes at the early PAS droplets with or without Atg8 lipidation reaction. Fluorescence images obtained with and without ATP were acquired on the same microscope under identical imaging settings (laser power, gain, and scan speed). This experiment was performed once. Scale bar = 10 μm. Yellow lines = 5.1, 4.3, 4.7, and 5.7 μm from left to right. b, FRAP analysis of R18-liposomes at early PAS droplets. Scale bar = 2 μm. Bars in the graph represent means ± s.d., n = 5 biological replicates. c, Observation of R18-containing liposomes with various concentrations of DOTAP at the early PAS droplets. Bottom shows the line profile along the yellow line. This experiment was performed once except for 50% DOTAP, which was performed three times with similar results. Scale bar = 10 μm. d, Effect of AIM WT or AIM mutant treatment on the interaction between GST-Atg8 and Atg1 analyzed by in vitro pulldown assay. This experiment was performed once. e, Observation of R18-containing liposomes with AIM WT or AIM mutant at the early PAS droplets. Bottom shows the line profile along the yellow line. This experiment was performed once. Scale bar = 10 μm.
