Figure 4. Hcy induces inflammasome assembly, leading to casp1 activation, and IL-1β cleavage in the presence/absence of LPS.

Celle were treated with L-Hcy (500µM)/LPS (10 µg/mL) as described in Fig. 1. Precursor and substrate of casp1, component of inflammasome and IL-1β were detected by IP and WB, respectively. A, Schematic sketch for Inflammasome complex assembly and cleavage. B, Inflammasome complex analysis. After Hcy/LPS treatment, cell lysate was used for IP and WB. C, IL-1β expression and activation. Cells were pretreated with Casp1 inhibitor for 30m prior to Hcy/LPS treatment. Protein levels were normalized by β-actin density. Numbers above on each bar is the percentage normalized by the mean of control. Arrows indicate the direction of significant changes. *, p < 0.05 vs control in same group; £, p < 0.05 vs Hcy treatment in same group; #, p < 0.05 vs same treatment in vehicle. LRRs: leucine-rich repeat; NACHT, acronym standing for NAIP (neuronal apoptosis inhibitor protein); PYD, pyrin domain; CARD, caspase activation and recruitment domain; P20, protein 20; P10, protein 10; Act-IL-1β, activated interleukin-1β. Values are Mean±SEM; n=4.