NLRP3 mediates NF-κB activation in sterile inflammation.

<p>(A and B) Real-time PCR analysis (A) and ELISA analysis (B) of TNF-α and IL-1β in shCtrl and shNLRP3 cell lines treated with 150 μg/ml MSU crystals for 90 min (A) or indicated period (B). (C) Immunoblot analysis of pro-IL-1β in cell lines treated with MSU crystals for the times indicated. (D) Real-time PCR analysis of TNF-α mRNA in cells treated with recombinant human IL-1β (1 ng/ml) or MSU crystals for 90 min. Values are normalized to an average of 1 in untreated cells. (E) Real-time PCR analysis of MyD88 (left panel) and TNF-α mRNA (middle and right panel) in negative-control siRNA-introduced (siCtrl) and MyD88 siRNA-introduced (siMyD#1, #2) cells left untreated (left panel) or treated with 10 ng/ml LPS (middle panel) or 150 μg/ml MSU crystals (right panel) for 60 min. (F) Real-time PCR analysis of IL-1β mRNA in shCtrl and shNLRP3 cell lines treated with 250 μg/ml aluminum adjuvants (Alm) for 140 min. (G) Real-time PCR analysis of NLRP3 (left panel) and IL-1β mRNA (right panel) in negative-control siRNA-introduced (siCtrl) and NLRP3 siRNA-introduced (siNLRP3) primary human monocytes treated with (right panel Alm +) or without (left panel, and right panel Alm −) 250 μg/ml of aluminum adjuvant for 140 min. All results are representative of at least three independent experiments. *<i>P</i><0.05, **<i>P</i><0.01.