Figure 6.

Triptolide inhibits NLRP3 inflammasome assembly in cardiac fibroblasts (CFs). (A) CFs were placed on glass slides and treated with AngII (1 μM) and/or triptolide (TP; 10 μg/mL) for 24 h. Cells were then stained for NLRP3 (Alexa Fluor 488; green) and ASC (Alexa Fluor 555; red) and observed under a laser confocal microscope; bar = 5 μm. (B) Experiments were performed as described in A, and the cells were stained for NLRP3 (Alexa Fluor 488; green) and caspase-1 (Alexa Fluor 555; red); bar = 5 μm. (C) Colocalization of NLRP3 with ASC or caspase-1, calculated using Pearson’s correlation coefficient (Rr) via ImageJ (n = 50). (D–E) Cells were transfected with a Flag-Nlrp3 construct and treated as in A. Total lysate was used for immunoprecipitation (IP) using an anti-Flag antibody, and the associated pro-caspase-1 and ASC were detected via immunoblotting (IB). Histograms represent the protein ratio normalized to Flag (n = 3) * p < 0.01 vs. medium; # p < 0.01 vs. AngII.