Fig. 5
Chloride efflux is essential for NLRP3 inflammasome activation. a–c ELISA of IL-1β in LPS-primed BMDMs which were transferred to a basic NaCl saline (130 mM NaCl, 5 mM KCl, 1 mM MgCl2, 1 mM CaCl2, 20 mM HEPES (pH 7.4), 1 mg/ml BSA, 10 mM glucose) or chloride-free NaGluc saline (130 mM NaGluc, 5 mM KGluc, 1 mM MgGluc2, 1 mM CaGluc2, 20 mM HEPES (pH 7.4), 1 mg/ml BSA, 10 mM glucose) and left stimulated with ATP a, MSU b, or nigericin c. d ELISA of IL-1β in LPS-primed BMDMs which were transferred to NaCl saline, NaBr saline (130 mM NaBr, 5 mM KBr, 1 mM MgBr2, 1 mM CaBr2, 20 mM HEPES (pH 7.4), 1 mg/ml BSA, 10 mM glucose), NaI saline (130 mM NaI, 5 mM KI, 1 mM MgI2, 1 mM CaI2, 20 mM HEPES (pH 7.4), 1 mg/ml BSA, 10 mM glucose), NaGluc saline or NaGlut (130 mM NaGlut, 5 mM KGlut, 1 mM MgGlut2, 1 mM CaGlut2, 20 mM HEPES (pH 7.4), 1 mg/ml BSA, 10 mM glucose) saline for different time points. e, f Immunoblot analysis IL-1β and cleaved caspase-1 e or ELISA of IL-1β in supernatants f of LPS-primed Nlrp3 +/+ or Nlrp3 –/– BMDMs which transferred to NaCl, NaBr or NaI saline for 3 h. g ELISA of IL-1β in supernatants of LPS-primed Clic4 –/– BMDMs transfected siRNA against Clic1 and Clic5 and left stimulated with nigericin for 30 min or transferred to NaCl, NaBr or NaI saline for 3 h. Data are from three independent experiments with biological duplicates in each (a–d, f, g; mean ± SEM of n = 6) or are representative of three independent experiments e. Student’s t-test a–c, f, g, ***P < 0.001
