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. 2018 Aug 6;217(8):2743–2763. doi: 10.1083/jcb.201710116

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© 2018 Gómez-Sánchez et al.

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Figure 4. — Atg2 requires PtdIns3P and lipid-packing defects to tightly associate with membranes in vitro. (A) GUVs with the same lipid composition as the liposomes used in Fig. 3 were incubated with either 400 nM purified Atg2-mGFP or an equal volume of buffer (control) for 5 min at room temperature before being imaged. Single focal plane (FP) images and maximum-intensity projections (MIPs) of 62 optical planes are shown. (B) Analysis of Atg2-mGFP binding to GUVs with different compositions. Where indicated, the lipid mixture used in A was altered by substituting 15 mol% DOPE(PE) with equal molarities of DOPC or ergosterol (erg), whereas 3 mol% PtdIns3P was replaced by an equal molarity of DOPC. Bars, 10 µm. (C) Quantification of Atg2-mGFP binding to GUVs of the experiment shown in B. At least 30 GUVs per sample were counted, and the graph represents means of three independent experiments ± SD.