FIG 8.

Effect of mitochondria on K1-CPS-mediated NLRP3 inflammasome activation. (A to C) J774A.1 macrophages were incubated for 30 min with or without cyclosporine (Cyclos. A), for 5.5 h with or without 1 μg/ml of K1-CPS, and then for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (A), IL-1β in the culture medium (B), and TNF-α in the culture medium (C) were measured by Western blotting and ELISA, respectively. (D and E) J774A.1 macrophages were incubated for 30 min with or without Mito-TEMPO, for 5.5 h with or without 1 μg/ml of K1-CPS, and then for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (D) and IL-1β in the culture medium (E) were measured by Western blotting and ELISA, respectively. In panels A and D, the results are representative of three separate experiments. In panels B, C, and E, the data are expressed as the means ± SD from three separate experiments. *, P < 0.05; *** P < 0.001.