Ab153724

The AMI and CAD tissues were lysed using strong RIPA buffer containing Halt Protease Inhibitor Cocktails (Thermo Fisher Scientific, Waltham, USA). Protein concentrations were evaluated with a bicinchoninic acid assay kit (Beyotime, Nantong, China). Primary antibodies targeting to beta actin (ab8226, Abcam), UHRF2 (ABE1028, MilliporeSigma), TET3 (ab153724, Abcam), UHRF2 (ZBTB33, MilliporeSigma), TET1 (ab19198, Abcam), DNMT3B (ab2851, Abcam), NTHL1 (ab191413, Abcam), UHRF1 (ab213223, Abcam), MBD3 (ab157464, Abcam), and SMUG1 (ab192240, Abcam), were incubated with targeted proteins at 4°C overnight, followed by incubating with appropriate horseradish peroxidase-conjugated secondary antibodies. Detection of horseradish peroxidase was performed with the Super Signal West Pico Chemiluminescent Substrate (Pierce).

The extraction of total protein from cells and tissue samples was performed by radio-imunoprecipitation assay lysis buffer with 1% phenylmethanesulfonyl fluoride, 1% phosphatase inhibitor, and 1% protease inhibitor (Beyotime Biotech, China). The experimental procedure was carried out as usual. The information of antibodies is listed as follows: anti-TET3 antibody (dilution, 1:1,000; ab153724; Abcam), anti-β-catenin antibody (dilution, 1:2,000; ab32572; Abcam), anti-p-AKT (T308) antibody (dilution, 1:1,000; ab38449; Abcam), anti-p-AKT (S473) antibody (dilution, 1:1,000; ab81283; Abcam), anti-p-GSK3β (Tyr216 + Tyr279) antibody (dilution, 1:2,000; ab68476; Abcam), anti-p-GSK3β (Ser9) antibody (dilution, 1:2,000; ab75814; Abcam), anti-GSK3β antibody (dilution, 1:5,000; ab32391; Abcam), anti-AKT antibody (dilution, 1:1,000; ab8805; Abcam), and anti-GAPDH antibody (dilution, 1:5,000; ab9482; Abcam).