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. 2020 Mar 10;11:352. doi: 10.3389/fimmu.2020.00352

Figure 5.

Figure 5

NLRP3 and caspase-1 specific activation of the inflammasome by NS2A and NS2B. HMEC-1 cells were transfected with expression 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 1 h as a positive control, in the presence or absence of Glyburide (200 μM). (A) The cell lysates were analyzed by western blot using an anti-Caspase 1 antibody (1:1,000), and (B) the cell free supernatant was analyzed for IL-1β by ELISA. (C) HMEC-1 cells were transfected with expression plasmids encoding NS2A-GFP, NS2B-GFP or the pEGFPN1 empty vector for 36 h or treated with LPS (1 μg/mL) for 6 h followed by ATP (5 mM) for 45 min as a positive control, in the presence or absence of AcVYAD-cmk (50 μM), and the cell free supernatant was analyzed for IL-1β by ELISA. Data are representative of at least three independent experiments and indicate the mean ± S.D. (B,C). ***P < 0.001.