FIGURE 5.

THIK‐1 specifically regulates ATP‐induced NLRP3 activation in bone‐marrow‐derived macrophages. (a) IL‐1β ELISA of the supernatant of primary wild‐type (WT) and Kcnk13 knockout (KO) BMDMs primed with LPS (1 μg ml⁻¹, 4 h) followed by pretreatment with MCC950 (10 μM) for 15 min before stimulation with ATP (5 mM, 1 h) (n = 7), silica (300 μg ml⁻¹,4 h) (n = 7), imiquimod (75 μM, 2 h) (n = 4) or nigericin (10 μM, 1 h) (n = 8). (b) Caspase‐1, IL‐1β and gasdermin D western blot of total cell lysates (cell lysate + supernatant) from LPS‐primed WT and Kcnk13 KO pBMDMs pretreated with vehicle control or MCC950 (10 μM) for 15 min before stimulated with ATP (5 mM, 1 h). (c and d) TNF and IL‐6 ELISA and NLRP3 and IL‐1β western blot of the supernatant and total cell lysates respectively of primary wild‐type (WT) and Kcnk13 knockout (KO) BMDMs pretreated with Bay11(10 μM) for 15 min before priming with LPS (1 μg ml⁻¹, 4 h) (n = 7). ****p < .0001, ***p < .001, **p < .01 determined by two‐way ANOVA with Bonferroni's post hoc analysis. Values shown are the mean ± SEM