Figure 2 YDZG inhibits NLRP3 expression. J774A.1 (A) or THP-1 (D) cells were pretreated with the indicated concentrations of YDZG for 6 h. NLRP3, caspase-1 and ASC expression was examined by Western blotting(n=5). (B) J774A.1 cells were treated or untreated with YDZG at 10 μM for different times. NLRP3, caspase-1 and ASC expression was examined by Western blotting (n=5). J774A.1 (C) and THP-1 (E) cells were pretreated with 10 μM YDZG 1 h before being stimulated with LPS (1 μg·mL-1) for another 6 h, and were then treated with ATP (5 mM) for 30 min before harvesting. NLRP3, caspase-1 and ASC expression was examined by Western blotting (n=5). (F) Immunoblot analysis of extracts from HEK293T cells transfected with NLRP3 plasmid (n=5). (G) HEK293T cells were transfected with eGFP-NLRP3 plasmid for 24 h. Cells were pretreated with YDZG (20 μM) for 1 h before being stimulated with LPS (1 μg·mL-1) for another 6 h, then treated with ATP (5 mM) for 30 min (n=3). Cells were fixed and stained with Hoechst 33342 (1 μM) for imaging (scale bar = 20 μm).