Quantitative real-time polymerase chain reaction (qRT-PCR)
Total RNA was isolated from 1×107 7-day-cultured ECs
using Trizol reagent (Life Technologies). qRT-PCR was carried out with
the
QuantiTect
SYBR Green RT-PCR kit
(Qiagen,
NRW, Germany) according to the manufacturer’s instructions. The relative
mRNA levels of TLR1, TLR2 and TLR4, CCR2, CCR5, CXCL10, PRF1 and the
NLRP3 inflammasome were analysed after normalization to 18s mRNA levels.