Fig. 3. NLRP3 inhibitor repressed KD-treated EC pyroptosis.

HUVECs were pretreated with NLRP3 inhibitor (MCC950, 10 μM) for 30 min, and then ECs were co-incubated with pooled sera-treated THP1 for 24 h. a Effect of MCC950 pretreatment on the protein expression of pyroptosis were analyzed by western blot analysis. GAPDH was used as an internal control. b–h Quantitative analysis of pyroptosis-related protein expression was conducted. Data were presented as mean ± SD (n = 3). i–k Caspase-1 expression, DNA fragmentation and LDH release were determined using immunofluorescent assay, TUNEL staining and LDH activity assay kits, respectively. Magnification: ×200, Scale bar = 100 μm. l Double staining of Hoechst 33342 (blue) and PI (red) was conducted in the treated endothelial cells (Left: the representative photographs, Right: the quantification evaluation of PI-positive cells). Magnification: ×100, Scale bar = 500 μm. Significance: *P < 0.05 and **P < 0.01 indicates significant difference between KD group and HC group, and ^#P < 0.05 indicates significant difference between KD groups with and without MCC950 addition