Anti β gal

All the antibodies employed in the present study are as follows: anti-β-gal (sc-19119, Santa Cruz, CA, USA), anti-GAPDH (sc-32233, Santa Cruz, CA, USA), anti-HDAC1 (c-19) (sc-6298, Santa Cruz, CA, USA), anti-HO-1 (H-105) (sc-10 789, Santa Cruz, CA, USA), anti-Nrf2 (ab89443, Abcam, Cambridge, UK), anti-p-Nrf2 (S40) (ab76026, Abcam), anti-PGC-1α (D-5) (sc-518025, Santa Cruz, CA, USA), anti-P16^INK4A (10883-1-AP, Proteintech, Manchester, UK), anti-p21 (F-5) (sc-6246, Santa Cruz, CA, USA), and anti-p53 (1C12) (no. 2524, Cell Signaling, Danvers, MA, USA), anti-SIRT1 (B-7) (sc-74465, Santa Cruz, CA, USA), and anti-β-actin (sc-47 778, Santa Cruz, CA, USA). The secondary antibodies, namely anti-rabbit, anti-mouse, and anti-goat, were purchased from Santa Cruz, California. H₂O₂, galangin, and resveratrol were purchased from Sigma-Aldrich (St. Louis, MO, USA).

The following antibodies were used in this study: anti-pJNK, anti-p-c-jun, anti-Bcl-xL, anti-β-gal, and anti-β-actin (Santa Cruz Biotechnology, Dallas, TX, USA). Anti-C-Caspase-3 was purchased from Cell Signalling Technology (Danvers, MA, USA). The ZAK monoclonal antibody (M02) was purchased from Abnova (Taipei, Taiwan). 3-HF was purchased from Sigma. siZAKβ was kindly provided by Dr. J. J. Yang (Chung Shan Medical University, Taichung, Taiwan).

Western blot analysis was conducted as previously described [31] (link). The nuclear/cytosol fractionation kit (Bio Vision Technology Inc., Canada) was used to separate nuclear and cytoplasmic proteins, according to the manufacturer's protocol. Proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to Protran nitrocellulose membranes (Whatman GmbH, Dassel, Germany). The signals were then detected with an Amersham ECL kit (GE Healthcare, Buckinghamshire, UK) and exposed to Amersham Hyperfilm ECL (GE Healthcare). The following antibodies were used: anti-Flag, (Sigma) anti-Smad3 [pSpS^423/425], anti-Smad2/3 (Invitrogen, Carlsbad, CA), anti-GAPDH (Epitomics, Burlingame, CA), anti-P450c17, anti-StAR, anti-3β-HSD, anti-Nur77, anti-β-gal, anti-α-Tubulin and anti-Lamin B (Santa Cruz Biotechnology, Santa Cruz, CA).