Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2022 Sep 7;609(7928):815–821. doi: 10.1038/s41586-022-05164-4

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s), under exclusive licence to Springer Nature Limited 2022, Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.

PMC Copyright notice

Fig. 4 — a, Live-cell imaging demonstrating rapid recruitment of eGFP–ATG2A to damaged lysosomes in U2OS cells stably expressing eGFP–ATG2A and LAMP1–mCherry. Scale bar, 10 μm. b, Quantification of eGFP–ATG2A puncta in individual U2OS cells upon LLOME treatment. c, eGFP–galectin-3 assay showing defects of rapid lysosomal repair in four independent clones of ATG2A/B-DKO cells. Data are mean ± s.e.m.; 50–100 cells, n = 3 for each condition. See more details in Extended Data Fig. 9a,b. d, Experimental design for the reconstitution of PS-stimulated lipid transport by ATG2A using a FRET-based assay. e, PS in acceptor liposomes potently stimulates ATG2A-dependent NBD fluorescence. Data are mean ± s.e.m.; n = 3 per condition. f, Illustration of lipid transport mutants of ATG2A. Flexible loops outside of the main structure are not shown. g, In vitro lipid transport assays testing the activity of various ATG2A mutants. Data are mean ± s.e.m.; n = 3 per condition. h, eGFP–galectin-3 assay demonstrating that ATG2A lipid transport mutants cannot rescue rapid lysosomal repair in ATG2A/B-DKO cells. Data are mean ± s.e.m.; 50–100 cells, n = 3 per condition. See fluorescence images in Extended Data Fig. 9f. i, In vitro ATG2A lipid transport assays in the presence of 1 μM maltose-binding protein (MBP) or MBP-tagged ATG2A-CT (MBP–CT). Data are mean ± s.e.m.; n = 3 per condition. j, Lysosomal repair defects in wild-type cells stably expressing LAMP1–CT but not in cells expressing LAMP1–mCherry or mCherry–CT. Data are mean ± s.e.m.; 50–100 cells were quantified, n = 3 per condition. See fluorescence images in Extended Data Fig. 9h. k, A cartwheel drawing of amino acids 1757–1789 from ATG2A-CT. Conserved residues mutated in the 5E mutant are indicated. l, The ATG2A(5E) mutant is not activated by PS-positive acceptor liposomes (A3) in lipid transport assays. Data are mean ± s.e.m.; n = 4. A2, charge-matched control acceptor liposomes. m, The ATG2A(5E) mutant cannot rescue rapid lysosomal repair in ATG2A/B-DKO cells. Data are mean ± s.e.m.; 50–100 cells were quantified, n = 3 per condition. See fluorescence images in Extended Data Fig. 9n. Unpaired, two-tailed t-tests.

Source data