Vegfr2 sirna

Some cohort cultures of HepG2 cultures were treated with histone H3 recombinant protein (20 μg/mL; Sigma‐Aldrich, Poole, UK), some cohort cultures were treated with VEGF recombinant protein (5 ng⁄mL; Thermo Fisher Scientific, Paisley, UK) or PBS vehicle for 24 hours, and some cohort cultures were treated with human nucleotide‐binding domain, leucine‐rich repeat containing protein 3 (NLRP3) siRNA (SI03060323; Qiagen, Crawley, West Sussex, UK), AIM2 siRNA (SI04261432; Qiagen), VEGF siRNA (sc‐29520; Santa Cruz Biotechnology, Dallas, TX), VEGFR1 siRNA (sc‐29319; Santa Cruz), VEGFR2 siRNA (sc‐29318; Santa Cruz), or scrambled siRNA (Qiagen) 6 hours before experiments. siRNA was dissolved in siRNA suspension buffer and administered to HepG2 cells at a dose of 20 nmol/L; scrambled siRNA served as a negative control. The transfection was carried out through highly efficient Lipofectamine Transfection Reagent (Thermo Fisher Scientific UK).21 Cells were incubated with siRNA for 6 hours before histone treatment. Some cell cohort received caspase‐1 inhibitor Ac‐YVAD‐CHO (20 μmol/L; Santa Cruz).

For transfection of VEGFR2 siRNA (Santa Cruz), HUVECs were cultured in 12-well plates. A transfection mixture of 100nM siRNA and 10 mg/ml lipofectin in serum-free medium was added to medium-aspirated cells for 4 h. The cells were then incubated with complete DMEM containing 10% FBS for 48 h before experiments.