A7028

According to the instruction manual of the ChIP kit (Cat. No. 492024, Thermo Fisher Scientific, Rockford, IL, USA), the SCC-9 or CAL27 cells were cross-linked in 1% formaldehyde and ultrasonicated to truncate DNA to 200-500 bp fragments. Thereafter, the lysates were reacted with the antibodies of NRIP1 (customized by Sangon Biotech), NSD2 (1:50, ab75359, Abcam), and H3K36me2 (1:50, ab176921, Abcam) overnight for immunoprecipitation, with normal rabbit IgG (A7016, Beyotime Biotechnology Co., Ltd., Shanghai, China) or normal mouse IgG (A7028, Beyotime) used as the NC antibodies. The DNA in the immunoprecipitated complexes was eluted and purified for qPCR analysis.

C57BL/6 J wild-type (WT) mice were purchased from the Jackson Laboratory (Sacramento, CA). Male mice (8 weeks old; n = 6 animals per group) were infused with saline or Ang II at a dose of 1000 ng/kg/min (Aladdin, CA) with an osmotic minipump (Alzet MODEL 1007D for 7 days; Alzet MODEL 1002 for 14 days; DURECT, Cupertino, CA) as previously described [3] . A control IgG (A7028, Beyotime, CA) or anti-ICAM-1 neutralizing antibody (BE0020-1, Bio X Cell, USA) was administered every 2 days beginning 1 day before the operation to block soluble ICAM-1 based on our preliminary results and previous publications [16] . The ICAM-1 neutralizing antibody was administrated by tail intravenous injection (IV.). All mice were maintained under specific pathogen-free conditions and provided free access to a normal diet (0.3% sodium chloride). After the study period, the mice were anesthetized, and the serum was collected and aortic tissues were removed and prepared for further molecular and histological examinations. All procedures were approved by the Institutional Animal Care and Use Committee of Dalian Medical University and conformed to the US National Institute of Health's Guide for the Care and Use of Laboratory Animals.