Rabbit anti nrf2

The cells were washed twice with cold PBS and lysed with RIPA buffer [50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1% NP-40, 0.25% sodium deoxycholate, 0.2 mg/ml leupeptin, 0.2 mg/ml aprotinin, 0.1 M phenylmethylsulfonylfluoride (PMSF), 1 mM Na₃VO₄ and 0.5 M NaF]. The lysates were centrifuged at 13,500 × g for 15 min at 4°C and the supernatants were loaded on to 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels. The following primary antibodies were used: rabbit anti-p53 (1:1,000; sc-6243; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA); rabbit anti-acetyl p53 (1:1,000; 06-758; Upstate Biotechnology, Lake Placid, NY, USA); mouse anti-p21 (1:2,000; sc-6246) and rabbit anti-Nrf2 (1:1,000; SC-722) (both from Santa Cruz Biotechnology, Inc.); mouse anti-α-tubulin (1:5,000; T5168; Sigma-Aldrich) and goat anti-lamin B (1:2,000; sc-6216; Santa Cruz Biotechnology, Inc.). Primary antibodies were detected using horseradish peroxidase-conjugated goat anti-mouse (A2554), -rabbit (A0545) (Sigma-Aldrich), or donkey anti-goat secondary antibodies (sc-2020; Santa Cruz Biotechnology, Inc.) and visualized using an enhanced chemiluminescence detection system (Thermo Fisher Scientific, Rockford, IL, USA).