FIG 2.

K1-CPS induces IL-1β secretion through the NLRP3 inflammasome. (A to C) J774A.1 macrophages were incubated for 30 min with or without the NLRP3 inhibitor glybenclamide (Glyb), for 5.5 h with or without 1 μg/ml of K1-CPS, and 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 to F) J774A.1 macrophages stably transfected with a control shRNA plasmid (sh-SC), an NLRP3 shRNA plasmid (sh-NLRP3), and an ASC shRNA plasmid (sh-ASC) were incubated for 5.5 h with or without 1 μg/ml of K1-CPS or LPS and then for 30 min with or without 5 mM ATP. The levels of activated caspase-1 (p10) in the cells (D), IL-1β in the culture medium (E), and TNF-α in the culture medium (F) 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, E, and F, the data are expressed as means ± SD from three separate experiments. ***, P < 0.001.