Fig. 3

Repeated NTG administration increased the expression of NLRP3 and IL-1β on microglia. a, b After repeated nitroglycerin (NTG) or saline administration, NLRP3 and IL-1β mRNA expression analysis were tested by qRT-PCR. Unpaired t test, ***p < 0.001 compared with the saline group (n = 6 mice per group). c, d The NLRP3 and IL-1β protein expression were tested by western blot on different days (NTG-0D, namely before the first injection of NTG). One-way ANOVA and Tukey’s multiple comparison test, *p < 0.05, **p < 0.01, ***p < 0.001 versus the NTG-0D group (n = 6 per mice group). e Double immunofluorescence staining for Iba-1 (red) with NLRP3 (green) in the mouse TNC after repeated NTG administrations. The white arrow shows the colocalization of Iba-1 and NLRP3 proteins. f Double immunofluorescence staining for Iba-1 (red) and IL-1β (green). The white arrow shows the colocalization of IL-1β and Iba-1 proteins. g Double immunofluorescence staining of IL-1β (red) and NLRP3 (green). The white arrow shows the colocalization of the IL-1β and NLRP3 protein. e–g × 40 objective lens; scale bars, 20 μm