Fig. 5
ZnONP-induced NLRP3 activation and pyroptosis are mediated by ROS in HaCaT cells. A Inhibitory effect of NAC on the ZnONP- and UVB-elicited production of ROS. B Histograms represent the fluorescence intensity of ROS. C Annexin V and PI were employed to examine ZnONP-induced HaCaT cell pyroptosis via flow cytometry. D Histograms represent the percentage of Q1 + Q2 regions, indicating PI positive cells. E Immunofluorescence staining with an anti-GSDMD antibody of HaCaT cells treated with UVB (68 mJ/cm2) + ZnONPs (10 μg/mL), UVB (68 mJ/cm2) + ZnONPs (10 μg/mL) + NAC (1 mM) for 24 h. Arrow indicate GSDMD puncta. F, G Western blot analysis of the effects of NAC on the ZnONP-induced NLRP3 inflammasome and pyroptosis proteins NLRP3, caspase-1, cleaved caspase-1, ASC, GSDMD and cleaved GSDMD-NT in HaCaT cells. H LDH release in ZnONP-treated HaCaT cells treated with NAC. Values are presented as the mean ± SD (n = 3). *p < 0.05, control group versus treatment groups. #p < 0.05, the UVB + ZnONPs groups versus the UVB + ZnONPs + NAC groups