Fig. 5: ATG9A loss impairs caveolae structure and sensitizes ovarian cancer cells to CAR-macrophage cytotoxicity.
From: Exploitable mechanisms of antibody and CAR mediated macrophage cytotoxicity
A Surface proteomic profiling comparing control (sgGAL4) and ATG9A knockout (sgATG9A) OVCAR-8 cells revealed significant downregulation of multiple caveolae-associated proteins, including CAVIN1 and CAVIN2 (highlighted with boxes). B Western blot validation confirming reduced expression of CAVIN1 and CAVIN2 upon ATG9A loss. C Quantification of CAVIN2-ATG9A colocalization in OVCAR-8 cells transfected with ATG9A-RFP and CAVIN2-GFP, either cultured alone or co-cultured with CAR macrophages for 24 h (n = 15 biological replicates, mean ± SEM). Statistical significance was determined using two-tailed unpaired Student’s t-tests. D Three-day IncuCyte co-culture experiment using control, ATG9A KO, CAVIN2 KO and CAVIN2 KO OVCAR-8 cells and α-EphA2-CAR macrophages (n = 3 biological replicates; mean ± SEM). Statistical significance was determined using two-tailed unpaired Student’s t-tests. E PI uptake assay measuring plasma membrane permeability OVCAR-8 cells infected with different sgRNAs co-cultured with α-EphA2-CAR macrophages (n = 5 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.
