Figure 4.

K_ATP channels and ABC transporters are dispensable for activation of the Cryopyrin inflammasome. (A–E) BMDMs from wild-type (WT) and SUR2^−/− (A and B), Kir6.1^−/− and Kir6.2^−/− (C and D), or abca1^−/−, abcg1^−/−, and abca1^−/−/abcg1^−/− (abca1/g1^−/−) mice (E) were left untreated (CTRL), stimulated with 10 µg/ml LPS for 3 h, treated with 5 mM ATP or 20 µM nigericin for 30 min, or stimulated with LPS and treated with ATP (LPS+ATP) or nigericin (LPS+nigericin). Cell extracts were immunoblotted for caspase-1 (A, C, and E), and culture supernatants were analyzed for secreted IL-1β (B and D). (F–H), LPS-primed BMDMs from abca1^−/−/abcg1^−/− (F), Kir6.1^−/− (G), and Kir6.2^−/− (H) mice were treated with 200 µM glyburide or DMSO for 15 min before 5 mM ATP or 20 µM nigericin was added for an additional 30 min. Cell extracts were immunoblotted for caspase-1. Black arrowheads indicate procaspase-1, and white arrowheads mark the p20 subunit. Cytokine data represent the mean ± SD of triplicate samples from a single experiment, and all results are representative of three independent experiments.