Fig. 7. Mitochondrial double-stranded RNA in exosome stimulates IL-1β production of human primary Kupffer cells.

(A) In EtOH-treated human hepatocytes, expression of genes (PNPT1 and SUPV3L1) and proteins (PNPase and SUV3) were assessed by qRT-PCR and Western blotting, respectively (12, 24 hour). Bar = 20 μm. (B) Representative immunostaining of J2 antibody in Veh- and EtOH (100 mM)-treated human primary hepatocyte (24 hour). dsRNA (Arrowhead), Bar = 10 μm. (C) Freshly isolated EtOH-EVs from primary human hepatocytes were treated to primary human Kupffer cells (hKC) for 24 hours and then gene expression was analyzed. (D) hKCs were co-incubated with WT γδ T cells (white arrowhead) for an additional 4 hours after treatments with Veh- and EtOH-Exo. (E) Primary hKC (black arrowhead) were treated with Veh- and EtOH-Exo in the absence and presence of RNase III (24 hour). (F) Primary hKC were treated with poly I:C (50 μg/g) for 3 hours and they were subjected to qRT-PCR. Values and images represent the results from three experimental replicates. Data are expressed as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 compared to the corresponding control, based on unpaired t-test between two groups and one way ANOVA with Dunnett’s test for multiple comparison vs control.