FIGURE 9.

Calcineurin inhibitors suppress IL-1β release during activation of the lipotoxic inflammasome. A–C, stimulated macrophages were incubated with the indicated concentrations of CSA, and IL-1β (A), TNFα (B), and LDH release (C) were quantified 20 h later. D and E, pMACs were incubated with vehicle (veh, red bars), CSA (black bars), or FK506 (FK, gray bars) during 20 h of stimulation with BSA-PBS or palmitate (palm)-LPS after which IL-1β (D) and TNFα (E) concentrations in the supernatant were determined by ELISA. F, mitochondrial membrane potential was assessed by tetramethylrhodamine ethyl ester (TMRE) staining followed by flow cytometry in pMACs incubated with vehicle (red line), CSA (black line), or FK506 (gray line) for 20 h. G–I, macrophages were stimulated in the presence of the indicated concentrations of CN585, and IL-1β (G), TNFα (H), and LDH release (I) were quantified 20 h later. Bar graphs indicate the mean ± S.E. for a minimum of three experiments, each performed in triplicate. *, p < 0.05 for vehicle versus inhibitor. ns, not significant.