Fig. 1

Nortriptyline triggers the inflammasome activation. a Nortriptyline structure. b and c Western blot of IL-1β and caspase-1 (b) and ELISA of IL-1β levels (c) in culture supernatants (SN) from BMDMs stimulated with nortriptyline in the absence or presence of LPS. d–h BMDMs were treated with LPS and nortriptyline. Western blot assessed the expression of mature caspase-1 and IL-1β in SN and GSDMD cleavage in whole-cell lysates (WCL) (d). The activity of caspase-1 (e) and the release of LDH (f) were assessed. The levels of IL-1β (g) and TNF-α (h) were evaluated by ELISA. (i–m) Cells were incubated with LPS and then treated with nortriptyline for 1, 3, 6, 12 h, respectively. Western blot evaluated the expression of mature IL-1β and cleaved caspase-1 in SN and GSDMD cleavage in WCL (i). The caspase-1 activity (j) and LDH (k) release in SN. ELISA assay the SN levels of IL-1β (l) or TNF-α (m). Data are expressed as the mean ± SEM (n = 3); *P < 0.05, **P < 0.01, ***P < 0.001 vs. the control group; ns, not significant; One-Way ANOVA analysis was followed by the Dunnett's post hoc test