Fig. 7: ATG9A-mediated plasma membrane repair after macrophage-induced damage is autophagy-independent.

A Western blot analysis of autophagic flux in control and KO OVCAR-8 cells. Chloroquine was added to prevent autophagosome degradation. B Western blot using control or sgATG9A SKOV-3/OVCAR-8 cells to examine p62 expression. C Overview of different roles of ATG9A. LC3: Microtubule-associated proteins 1 A/1B light chain 3; PE: Phosphatidylethanolamine; SNX18: Sorting Nexin 18; AP-1: Adapter Protein Complex 1; AP-2: Adapter Protein Complex 2; Rab11: Ras-related protein Rab-11A; VAMP7: Vesicle-associated membrane protein 7; Syt VII: Synaptotagmin-7; SNAP-23: Synaptosomal-associated protein, 23 kDa; STX-4: Syntaxin-4; TGN: Trans-Golgi Network; Faa1: Fatty acid amide hydrolase 1; ER: Endoplasmic Reticulum. D Competition assay using regular media or media with Autophinib (n = 3 biological replicates; mean ± SEM). Statistical significance was determined using two-tailed unpaired Student’s t-tests. E CAR-M Co-culture experiments using sgGAL4, sgATG9A OVCAR-8 cells and sgATG9A cells with Autophinib treatment (n = 4 biological replicates; mean ± SEM). Statistical significance was determined using two-way ANOVA with Tukey’s multiple comparisons test. F-G Live cell percentage or relative PI+ number were normalized to the cell count or PI+ cell number in the cancer-only group (n = 4 biological replicates; mean ± SEM). Statistical significance was determined using two-tailed unpaired Student’s t-tests. H Cell proliferation at day 3 normalized to cell number at day 0 (n = 3 biological replicates; mean ± SEM). Statistical significance was determined using two-tailed unpaired Student’s t-tests. *P < 0.05; **P < 0.01; ***P < 0.001. Some elements of this figure were created with BioRender.com and are included under a publication license in accordance with BioRender’s user agreement. Created in BioRender. DeSelm, C. (2025) https://BioRender.com/xrev1ob.