Fig. 6
Clics act downstream of potassium efflux or mitochondrial damage to promote NLRP3 activation. a, b Qualification of the decrease of intracellular potassium in LPS-primed Clic4 –/– BMDMs transfected with siRNA against Clic1 and Clic5 a or Nlrp3 –/– BMDMs tranfected with siRNA against Clic1, Clic4 and Clic5 b and left stimulated with ATP or nigericin. c ELISA of IL-1β in supernatants of LPS-primed Clic4 –/– BMDMs transfected siRNA against Clic1 and Clic5 and left transferred to potassium-free saline (135 mM NaCl, 1 mM MgCl2, 1 mM CaCl2, 20 mM HEPES (pH 7.4), 1 mg/ml BSA, 10 mM glucose) for 3 h. d Confocal microscopy analysis of LPS-primed Clic4 –/– BMDMs transfected with siRNA against Clic1 and Clic5 and then left stimulated with nigericin, followed by staining with Mitotracker red and DAPI. e ELISA of IL-1β in supernatants of LPS-primed BMDMs transfected stimulated with nigericin or transferred to NaBr, NaI saline or potassium-free saline with or without the presence of indicated doses of MnTBAP. Data are from three independent experiments with biological duplicates in each (a–c, e; mean ± SEM of n = 6) or are representative of three independent experiments d. Two-way ANOVA e, Student’s t-test a–c, ***P < 0.001, NS not significant
