Anti hnrnp c1 c2 sc 32308

Cells were washed with ice-cold phosphate buffer saline and lysed in RIPA buffer (Thermo Fisher Scientific), supplemented with a complete protease inhibitor cocktail (Roche, Basel, Switzerland). Equal protein amounts were resolved in 10% Bis–Tris Plus Gels (Thermo Fisher Scientific), transferred onto the BioTrace NT pure nitrocellulose blotting membrane (PALL Corporation) and blocked with 5% non-fat dry milk in Tris-buffered saline pH 7.6 containing 0.1% Tween-20. Proteins were probed with corresponding antibodies (listed below) and detection was performed using WestDura (Thermo Fisher Scientific) with myECL Imager (Thermo Fisher Scientific). Antibodies: anti-p53 rabbit pAbs (CM-1-Recamo); anti-actin mouse pAbs (AC-15, Sigma-Aldrich), anti-hnRNP C1/C2 (sc-32308, Santa Cruz), anti-PTB (32–4800, Thermo Fisher Scientific), anti-eIF2α (sc-133132, Santa Cruz) HRP-conjugated secondary antibodies (Dako, Glostrup, Denmark). Western blots represent n ≥ 3 and the original uncropped blots are provided in the Supplementary Information.

Human SMC growth medium (M231), smooth muscle growth supplement, trypsin/ethylenediaminetetraacetic acid (EDTA) solution, trypsin neutralizer solution, and HASMCs were obtained from Cascade Biologics (Portland, OR). Fetal bovine serum (FBS), antibiotic–antimycotic mixture, mouse TNF-α kits, oligofectamine, and 2’, 7’-dichlorofluorescein diacetate were obtained from Life Technology (Grand Island, NY). Small interfering RNA (siRNA) oligonucleotides against RAGE and antibodies against human TNFR-1 (sc-8436), MSX2 (sc-17729), BMP-2 (sc-6895), RUNX2 (sc-10758), CD68 (sc-9139), β-actin (sc-47778), and anti-hnRNP c1/c2 (sc-32308) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Antibodies against human Nox subunit p47 (#610354), and caveoli-1 (#610406) were obtained from BD Biosciences (San Jose, CA). Unless otherwise specified, all other chemicals and reagents obtained from Sigma-Aldrich (St. Louis, MO). All methods in this study were reported in accordance with ARRIVE guidelines.