Anti acetyl p53 lys379

For total protein extracts, tissues were homogenized in protein lysis buffer and centrifuged at 15,000g for 15 minutes at 4ºC. Nuclear extracts were isolated as previously described.^{57 (link)} Proteins were separated by 8% or 10% wt/vol sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE), transferred to polyvinylidenedifluoride (PVDF) membranes and probed overnight at 4°C. The primary antibodies used were: Anti-Sirt1 (07-131, Millipore, Billerica, MA), Anti-AMPK (2532, Cell Signaling, Danvers, MA), Anti-phospho-AMPK-Thr¹⁷² (2531, Cell Signaling), Anti-AKT (9272, Cell Signaling), Anti-phospho-AKT-Ser⁴⁷³ (9271, Cell Signaling), Anti-Acetyl-p53-Lys³⁷⁹ (2570, Cell Signaling), Anti-Myosin Skeletal Slow (M8421, Sigma-Aldrich, St. Louis, MO), Anti-p53 (2524, Cell Signaling) and Anti-α-tubulin (ab4074, Abcam, Cambridge, UK). Detection was performed using the corresponding horseradish peroxidase-labeled secondary antibodies and western blotting detection reagent (ECL Plus; Amersham, Freiburg, Germany).

For total protein extracts, tissues were homogenized in protein lysis buffer and centrifuged at 15,000g during 15 minutes at 4 °C. Nuclear extracts were isolated as previously described.^{24 (link)} The primary antibodies used were: Anti-Sirt1 (Millipore 07-131, Billerica, MA), Anti-phospho-ACC-Ser⁷⁹ (Millipore 07-303), Anti-AMPK (Cell Signalling 2532, Danvers, MA), Anti-phospho-AMPK-Thr¹⁷² (Cell Signalling 2531), Anti-phospho-STAT3-Ser⁷²⁷ (Cell Signalling 9134), Anti-ERK1/2 (Cell Signalling 9102), Anti-phospho-ERK1/2-Thr²⁰²/Tyr²⁰⁴ (Cell Signalling 9101), Anti-Acetyl-p53-Lys379 (Cell Signalling 2570), Anti-PGC1 (Santa Cruz H-300, Dallas, TX), Anti-α-tubulin (Abcam ab4074, Cambridge, UK). Detection was performed using the corresponding horseradish peroxidase-labeled secondary antibodies and Western blotting detection reagent (ECL Plus; Amersham, Freiburg, Germany).

DEHP (#80030) and flutamide (#F9397) were purchased from Sigma-Aldrich. Primary antibodies used for immune blot were: anti-Caspase-8 (#9746), anti-Caspase-9 (#9502), anti-Caspase-3 (#9662), anti-PARP-1 (#9542), anti-Bcl-xL (#2764), anti-Bid (#2003), anti-Bax (#2772), anti-Bak (#6947), anti-acetyl p53 (Lys379, #2570) and anti-SIRT1 (#8469) antibodies from Cell signaling; Anti-p53 (#sc-71818) was from Santz Cruz; anti-actin (##ab6276) and mitoProfile total OXPHOS rodent Wb antibody cocktail (#ab110411) were from Abcam. All other reagents were from Sigma-Aldrich unless otherwise indicated.