Anti acetylation

Western blotting was performed as previously described [16 (link)]. The primary antibodies used were as follows: anti-cleaved-caspase-3, anti-Bcl-2, anti-Bax, anti-phosphorylated-AMPK, anti-AMPK, anti-phosphorylated-Akt, anti-Akt, anti-LC3B, anti-p62, anti-Beclin1, anti-ATG5, anti-ATG7, anti-Acetylation, anti-Lamin B and anti-SIRT1 antibody from Cell Signaling Technology, Inc. (Danvers, MA). The anti-β-tubulin1 antibody was from Boster Biological Technology (Wuhan, China). The secondary antibodies were provided by Boster Biological Technology.

NaB and SAHA were purchased from Sigma Aldrich (Deisenhofen, Germany). The FOXO1 inhibitor AS1842856 (AS) was obtained from Selleck, and the adenosine monophosphate-activated protein kinase (AMPK) inhibitor was purchased from MCE. The monoclonal anti-ubiquitin, anti-acetylation, anti-Snail, anti-Smad2, anti-Smad3, anti-Smad2/3, anti-p-Smad2/3, anti-FOXO1, anti-LC3B, anti-P62, anti-ULK1, anti-p-ULK1, anti-LC3, anti-GAPDH, and anti-vimentin antibodies and the secondary anti-mouse or anti-rabbit antibody conjugated to horseradish peroxidase (HRP) were acquired from Cell Signaling Technology (MA, USA). The secondary anti-rabbit antibody conjugated to fluorescein isothiocyanate (FITC), Protein A/G Sepharose and anti-AMPK were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-p-AMPK and anti-FN were acquired from Abcam. A Millicell chamber (8 mm) was purchased from Millipore (BD Biosciences, USA). The Annexin V, FITC apoptosis detection Kit S was obtained from Japan Tongren Chemical. SYBR Premix ExTaq II was a product of TaKaRa (TBI, Japan). siRNA FOXO1 and siRNA ULK1 were purchased from RIBO, and Lipofectamine 3000 was purchased from Invitrogen (Carlsbad, CA, USA). 4,6-diamidino-2-phenylindole (DAPI) dye was obtained from FUDE company.