Effects of GW-A2 on NLRP3 inflammasome activation.

<p>(<b>A</b> and <b>B</b>) J774A.1 macrophages were incubated for 30 min with or without 2 μM of GW-A2, for 5.5 h with or without 0.1 μg/ml of <i>E</i>. <i>coli</i> LPS, and then for 30 min with or without 5 mM of ATP (<b>A</b>) or 10 μM of nigericin (<b>B</b>). The levels of IL-1β in the culture medium and activated caspase-1 (p10) in the cells were measured by ELISA and Western blotting, respectively. (<b>C</b>—<b>F</b>) J774A.1 macrophages were incubated for 5.5 h with 0.1 μg/ml of <i>E</i>. <i>coli</i> LPS, for 30 min with or without 2 μM of GW-A2, and then for 30 min with or without 5 mM of ATP (<b>C</b> and <b>E</b>), 10 μM of nigericin (<b>D</b>) or 100 μg/ml of MSU (<b>F</b>). The levels of IL-1β and TNF-α in the culture medium and activated caspase-1 (p10) in the cells were measured by ELISA and Western blotting, respectively. The data are expressed as the mean ± SD of three independent experiments. The Western blotting results shown are a representative experiment of three independent experiments. *, ** and *** indicate significant differences, representing <i>p</i> < 0.05, <i>p</i> < 0.01 and <i>p</i> < 0.001, respectively compared to stimulation alone.