Figure 1.
Faa1 binds Atg9 vesicles as well as artificially generated Atg9 proteoliposomes. (A) Model for phagophore expansion driven by localized de novo phospholipid synthesis. (B) Microscopy-based membrane binding assay testing the recruitment of Faa1-mCherry to isolated and immobilized endogenous Atg9 vesicles with and without active PI3KC3C1. The experimental setup is shown on the left. (C) Microscopy assay testing the recruitment of Faa1-mCherry to ATTO 390-1,2-dioleoylphosphatedylethanolamine (DOPE) labeled Atg9 proteoliposomes. Atg9-proteoliposomes were immobilized on GFP-trap beads. Quantification of Faa1 recruitment to Atg9 proteoliposomes can be seen on the bottom left. Data are means ± SD (n = 3). The lipid composition of the Atg9 vesicle-like proteoliposomes is 42.5% POPC, 6% POPS, 6% POPE, 41.5% liver PI, 2.5% PI3P, and 1.5% ATTO 390-DOPE. PL = proteoliposome.

Faa1 binds Atg9 vesicles as well as artificially generated Atg9 proteoliposomes. (A) Model for phagophore expansion driven by localized de novo phospholipid synthesis. (B) Microscopy-based membrane binding assay testing the recruitment of Faa1-mCherry to isolated and immobilized endogenous Atg9 vesicles with and without active PI3KC3C1. The experimental setup is shown on the left. (C) Microscopy assay testing the recruitment of Faa1-mCherry to ATTO 390-1,2-dioleoylphosphatedylethanolamine (DOPE) labeled Atg9 proteoliposomes. Atg9-proteoliposomes were immobilized on GFP-trap beads. Quantification of Faa1 recruitment to Atg9 proteoliposomes can be seen on the bottom left. Data are means ± SD (n = 3). The lipid composition of the Atg9 vesicle-like proteoliposomes is 42.5% POPC, 6% POPS, 6% POPE, 41.5% liver PI, 2.5% PI3P, and 1.5% ATTO 390-DOPE. PL = proteoliposome.

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