Fig. 2: Fluoxetine triggers the maturation of downstream effector cytokines and cleavage of GSDMD.

A The fluoxetine chemical structure. B–F BMDMs were pretreated with LPS and subsequently stimulated with a range of fluoxetine concentrations. The cleaved caspase-1, as well as IL-1β in SN and the cleaved GSDMD in WCL (B) were assessed by western blotting. The caspase-1 activity (C) and the LDH release (D) were measured. Using ELISA to measure the secretion of IL-1β (E) and accumulation of TNF-α (F) in SN. G–K BMDMs were first primed with LPS and then stimulated with fluoxetine (40 μM) for 1, 3, 6, and 12 h, respectively. The expression levels of the cleaved caspase-1 and IL-1β in SN and the cleaved GSDMD (G) in WCL were detected using western blot analysis. Activity of caspase-1 (H), release of LDH (I), secretion of IL-1β (J) as well as generation of TNF-α (K) in SN were also assessed. L PMA-primed THP-1 cells were stimulated with fluoxetine and MSU, and then the expression levels of cleaved caspase-1 and IL-1β were detected by western blotting. Data are represented as the mean ± SEM from three biological samples, *P < 0.05, **P < 0.01, ***P < 0.001 vs. the control, ns, no significant. One-Way ANOVA analysis was followed by Dunnett’s post-hoc test.