FIGURE 4.

DHT has no effect on upstream signaling events of NLRP3 inflammasome activation and NLRP3–NEK7 interaction. (A) Qualification of potassium efflux in LPS-primed BMDMs exposed to various doses of DHT (2.5, 5, 10 μM) and then stimulated with nigericin. (B) ATP-induced Ca²⁺ flux in LPS-induced BMDMs treated with or without DHT was measured using a FLIPR Tetra system. (C) LPS-primed BMDMs treated with DHT and then stimulated with nigericin were detected by staining with MitoSox. The percentage of ROS-positive cells was obtained by flow cytometry. (D) HEK-293T cells were transfected with Flag-NLRP3, and DHT (10 μM) was added at 18 h post transfection. Immunoprecipitation was performed with anti-Flag affinity gel-agarose beads, followed by Western blot analysis. GAPDH was used as the lysate loading control. Data are shown as mean ± SEM from three biological replicates. One-way ANOVA was used to analyze the data. *p < 0.05, **p < 0.01, ***p < 0.001, NS: not significant.