TRIM5 protects cells from excessive inflammation and cell death in response to mitochondrial damage
(A–D) Seahorse analysis of oxygen consumption rate (OCR) in WT and TRIM5 knockout Huh7 cells in response to IVM (1.5 μM) or DMSO alone. n = 3 biological replicates.
(E)Immunoblot analysis of immune- and inflammation-related proteins in WT or TRIM5 knockout Huh7 cells following a 24 h treatment with 15 μM IVM or DMSO control. Plots show the abundance of the indicated protein relative to a loading control (actin). n = 3 biological replicates.
(F)The impact of IVM treatment (20 μM; 24 h) on the viability of WT and TRIM5 knockout Huh7 cells. The intensity of propidium-iodide labeling, indicative of plasma-membrane disruption, was determined in single cells by flow cytometry.
(G)Schematic illustration of TRIM5 action in mitophagy. Left: a portion of the cellular pool of TRIM5 associates with mitochondria under basal conditions.Following mitochondrial damage, TRIM5 links mitophagy eat-me tags (e.g., NIPSNAP2) with FIP200, ATG13, and other autophagy proteins (ATGs) at ER-mito-chondria contact sites. Data: mean + SEM; p values determined by ANOVA; *p < 0.05, **p < 0.01, ***p < 0.001, †, not significant.