Figure 5.

EMR2 ligation triggers the activation signal for NLRP3 inflammasome activation in human primary monocytes. (A–F) Isolated CD14⁺ primary monocytes (2 x 10⁶ cells/well) were incubated in 6-well plates coated without or with 2A1 mAb in the absence or presence of LPS for 24 h (A–D). When necessary, cells were pretreated with Ac-YVAD-CHO (50 μM) for 1 h before treatment with LPS and 2A1 (C, D) or cultured in medium containing 5% NHS or ATP (5 mM) as described (E, F). Cell supernatants were collected and analyzed for IL-1β by ELISA (A, C, E, F). Cell lysates were analyzed by WB analysis for relevant proteins as indicated (B, D). Data are means ± SEM of three independent experiments performed in triplicate using cells from three different donors (E, F). Data were analyzed by one-way ANOVA. ^* p<0.05, ^** p<0.01, ^*** p< 0.001, ^**** p<0.0001 versus the control group. ns, non significant.