Fig. 10.

NLRP3 inflammasome activation in microglia cells exposed to ER stress. Protein levels of GRP78 (A) were quantified by WB in total extracts obtained after treatment of BV2 cells with 2 or 10 μg/mL brefeldin A (BFA) for 6 h. NLRP3 (B) and pro-IL-1β (C) content was also quantified by WB in total extracts of BFA-treated BV2 cells upon LPS priming (3 h). Actin was used as a protein loading control to normalize the levels of the protein of interest. Results were calculated relatively to control values and represent the mean ± SEM of at least three independent experiments. Statistical significance between control and BFA-treated cells and between control and LPS plus BFA cells was determined using the one-way ANOVA test, followed by the Dunnett’s post hoc test: *p < 0.05, **p < 0.01, ***p < 0.001. Secreted IL-1β levels (D) were measured using an ELISA assay in the supernatants collected from BV2 microglia cells treated with 300 ng/mL LPS alone (3 h), or primed with LPS and then incubated with BFA 2 or 10 μg/mL) for 6 h. Results represent the mean ± SEM of at least three independent experiments. Statistical significance between LPS and control conditions (Ctrl), in the absence of treatments, was determined by student’s t-test (^##p < 0.01), and between LPS and LPS plus BFA cells was determined using the one-way ANOVA test, followed by the Dunnett’s post hoc test: *p < 0.05