Figure 4

Canonical and non-canonical release of inflammatory mediators and DAMPs is regulated by NLRP3. BEAS-2B cells were transfected with control (siControl) or NLRP3 siRNA (siNLRP3). Experiments were performed 24 h after siRNA transfection. A. Knockdown of NLRP3 mRNA levels by siRNA. B. Western blot analysis of whole cell lysates for the level of NLRP3 expression. Actin was used as a loading control. ELISA was performed on concentrated medium SN for detection of secreted IL-1β (C) and bFGF (D) 24 h post-transfection. The effect of LPS priming (5 μg/mL) was also investigated. E. Western blot analysis, to detect influence of NLRP3 knockdown on HMGB1 levels was conducted on concentrated medium supernatants. Histogram represents semi-quantitative densitometric analysis of HMGB1 expressed as arbitrary units. Data are presented as mean fold change ± SEM with *p-value <0.05 compared to UC, and # p-value <0.05 compared to the same treatment in the siControl setting.