FIG. 1.
tBHQ inhibits NLRP3 inflammasome activation. THP-Ms were treated with tBHQ for 12 h, and the protein level of Nrf2 and its nuclear translocation were determined by Western blot (A). tBHQ-pretreated THP-Ms were stimulated with LPS for 4 h and ATP for 1 h. The panel shows a Western blot of NLRP3, cleaved caspase-1 (p10), and cleaved IL-1β (p17) in the supernatants or cell lysates (B). IL-1β was measured by ELISA (C). The mRNA levels of NLRP3 and pro-IL-1β were detected by real-time RT-PCR (D, E). Western blot analysis of Nrf2 expression in THP-Ms transfected with different concentrations of Nrf2 siRNA or Nrf2 plasmid for 12 h (F). NLRP3, cleaved caspase-1, and IL-1β were detected by Western blot (G). Nrf2 siRNA-transfected THP-Ms were stimulated with LPS and ATP for the indicated periods. NLRP3, cleaved caspase-1, and IL-1β were detected by Western blot (H). IL-1β was measured by ELISA (J). Nrf2 plasmid-transfected THP-Ms were stimulated with LPS and ATP for the indicated periods. NLRP3, cleaved caspase-1, and IL-1β were detected by Western blot (I). IL-1β was detected by ELISA (K). The results are representative of three independent experiments and are expressed as the mean ± SD. *p < 0.05, **p < 0.01 compared with the control, #p < 0.05, ##p < 0.01 compared with LPS + ATP. ATP, adenosine triphosphate; LPS, lipopolysaccharide; NLRP3, NLR family, pyrin domain containing 3; Nrf2, nuclear factor erythroid 2-related factor 2; tBHQ, tert-butylhydroquinone; THP-Ms, THP-1 cell-derived macrophages.