Fig. 8.

NLRP3 inflammasome activation in human monocytes exposed to ER stress. Protein levels of NLRP3 (A), pro-IL-1β (B) and ASC (C) were quantified by WB in total extracts from THP-1 monocytes incubated in the presence or absence of 10 μg/mL TM for 8 h, upon priming with LPS (1 μg/mL) during 24 h. β-Tubulin I was used as a loading control to normalize the levels of the protein of interest. Results were calculated relatively to control values, with exception of pro-IL-1β, and represent the mean ± SEM of at least three independent experiments. Statistical significance between control and TM-treated cells was determined using the one-way ANOVA test, followed by Dunnett’s post hoc test: ****p < 0.0001. Levels of secreted IL-1β were quantified by an ELISA assay in supernatants of THP-1 cells treated with 1 μg/mL LPS alone (24 h), or with LPS (24 h) and then with 10 μg/mL TM for the last 8 h. Cells primed with 1 μg/mL LPS for 24 h and then exposed to 5 μM ATP for 30 min were used as a positive control for NLRP3 activation. Results were calculated relatively to LPS-treated cells and 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 < 0.0001), and between LPS, LPS plus ATP and LPS plus TM was determined using the one-way ANOVA test, followed by Dunnett’s post hoc test: *p < 0.05