Fig. 7

Exosomes containing NLRP3, ASC and Caspase-1 proteins were increased when THP-1-MØ cells were exposed to ZnONPs. Exo-ZnONPs from THP-1-MØ triggered NLRP3 inflammasome-related protein and proinflammatory cytokine expression in HaCaT cells. A Characterization of THP-1-derived macrophage-secreted exosomes. Exosomes were isolated from the macrophage culture medium, and exosome markers, including CD63, TSG101, and Flotillin-1, were identified by western blotting. The number of particles isolated from the culture medium was determined by NTA. B Representative TEM images of exosomes obtained from untreated macrophage culture medium. Scale bars: 100 nm. C THP-1-derived macrophages were exposed to ZnONPs (10 and 15 μg/ml) for 16 or 24 h. The exosomes in the different groups were extracted, and the exosome cargo proteins NLRP3, ASC, and Pro-caspase-1 were assessed by western blotting. Flotillin-1 was used as a loading control. Histograms representing the quantification of NLRP3, ASC, and pro-caspase-1 normalized to flotillin-1 in three independent experiments. The results were expressed as the mean ± SD. D HaCaT cells were incubated with macrophage-derived Exo-control, Exo-ZnONPs, and Exo-ZnONPs + Triton 0.1% for 24 h. HaCaT cells were extracted, and the expression of the NLRP3 inflammasome-related proteins TNF-α and IL-1β was assessed by western blotting. GAPDH was used as a loading control. E Histograms represent the quantification of NLRP3, ASC, caspase-1, caspase-1(p20), TNF-α, and IL-1β normalized to GAPDH in three independent experiments. The results are expressed as the mean ± SD. (*p < 0.05, **p < 0.01, ***p < 0.001 compared with control)