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. Author manuscript; available in PMC: 2014 Aug 6.
Published in final edited form as: Circulation. 2013 Jul 9;128(6):632–642. doi: 10.1161/CIRCULATIONAHA.113.002714

Figure 4.

Figure 4

SREBP2 Overexpression Up-regulates NLRP3 Inflammasome. (A) Depiction of the putative SRE binding site located at −1379/−1368 bp upstream of the transcription initiation site in the human NLRP3 promoter. The NLRP3 mRNA levels (B) and representative immunoblot (and quantification) of NLRP3 and the mature form of SREBP2 (C) in HUVECs treated with Ad-null (5MOI), Ad-SREBP2(N) (2.5 MOI or 5 MOI). (D) NLRP3 mRNA levels in lung ECs from wild-type or EC-SREBP2(N)-Tg mice (n=8). (E) ChIP analysis with antibodies against SREBP2 or IgG, soluble chromatin from HUVECs infected with Ad-null or Ad-SREBP2(N), and primers targeting the region spanning the SRE binding site in the NLRP3 promoter. (F) Representative immunoblot of NLRP3, precursor SREBP2, and mature form of SREBP2 in HUVECs transfected with 20 nM control RNA or SREBP2 siRNA for 48 hr, then exposed to static conditions or OS for 14 hr. Bar graphs represent mean ± SEM from at least 3 independent experiments. Mann-Whitney U test with exact method was used. ‘*’ indicates p < 0.05 vs Ad-null or wild-type whereas ‘#’ indicates p = 0.05 vs Ad-null.