GEF-H1 is not required for NLRP3-mediated IL-1β secretion, capase-1 processing, and LDH release in LPS-primed macrophages incubated with nigericin. (A) Quantitative RT-PCR analysis of Nlrp3, Pycard, and Casp1 mRNA expression in untreated wild-type (WT) and Arhgef2-/- bone marrow macrophages. Values are normalized to the expression of Gapdh. (B) Representative western blot analysis of NLRP3, ASC, and Pro-Caspase-1 (ProCasp1) in untreated WT and Arhgef2-/- (-/-) BMDMs. (C) IL-1β released by LPS-primed WT and Arhgef2-/- BMDMs treated with NLRP3, AIM, or NLRC4 inflammasome inducers. (D) Representative western blot analysis of pro-caspase-1 (Casp1 p45), active-caspase-1 subunit p20 (Casp1 p20), and ASC in the supernatants or cell lysates of LPS-primed WT and Arhgef2-/- (-/-) BMDMs left unstimulated or incubated with indicated inflammasome activators. (E) LDH released by LPS-primed WT and GEF-H1-deficient BMDMs incubated with nigericin. n = 3 independent experiments per group. Data represent the mean ± SD. Statistical analyses were performed using an unpaired two-tailed t-test.