Figure 6.

Requirement of increased intracellular Ca⁺⁺ concentration for NLRP3 inflammasome activation by NS2A and NS2B. HMEC-1 cells were transfected with plasmids encoding NS2A-GFP, NS2B-GFP, or the pEGFPN1 empty vector for 36 h or treated with LPS (1 μg/mL) for 6 hrs followed by ATP (5 mM) for 45 min as a positive control, in the presence or absence of BAPTA-AM (10 μM). (A) The cell lysates were analyzed by western blot using an anti-caspase-1 antibody (1:1,000), and (B) the cell free supernatants were analyzed for IL-1β by ELISA. Data are representative of at least three independent experiments, and indicate the mean ± S.D. (B). ^*P < 0.05 and ^***P < 0.001. ns, non significant.