Transdux transduction reagent

Antibodies against human SIRT1 (#2310), SIRT3 (#2627), PGC-1α (#2178) and acetylated-lysine (#9441) were all purchased from Cell Signaling Technology, Inc. (Beverly, MA, USA). The antibody against human CyPD (#ab110324) and mtTFA Human SimpleStep ELISA™ kit were purchased from Abcam, Inc. (Cambridge, MA, USA). The SIRT3 Deacetylase Fluorometric assay kit was purch ased from CycLex Co., Ltd. (Nagano, Japan). SIRT1 open reading frame (ORF) and control vector were both obtained from the Functional Genomics Facility of the University of Colorado. Lipofectamine 2000 was purchased from Life Technologies, Inc. (Grand Island, NY, USA). TransDux™ transduction reagent was purchased from System Biosciences, Inc. (Mountain View, CA, USA). Human Mitochondrial to Nuclear DNA Ratio kit was purchased from Clontech Laboratories, Inc. (Mountain View, CA, USA). CellTiter-Glo Luciferase-based assay was purchased from Promega (Madison, WI, USA). Specific siRNA for human SIRT3 and PGC-1α and scrambled siRNA were purchased from Integrated DNA Technologies, Inc. (Coralville, IA, USA). HiPerFect^® transfection reagent and other transfection-related reagents were purchased from Qiagen (Valencia, CA, USA). Dulbecco's modified Eagle's medium (DMEM) was purchased from Life Technologies. All other chemicals and reagents were from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA).

Antibody against human Osx was purchased from Santa Cruz Biotechnology, Inc. Antibodies against human α‐SMA were purchased from Abcam, Inc. Antibodies against human Runx2, vimentin, phosphatase and tensin homolog (PTEN), phosphorylated protein kinase B (AKT), and total AKT were purchased from Cell Signaling, Inc. The miR mimics and antagomirs, control miR, HiPerFect transfection reagent, other transfection‐related reagents, and the EndoFree Plasmid Maxi Kit were purchased from Qiagen. Lentivirus vector expressing miR mimic or antagomir and BLOCK‐iT Lentiviral Pol II miR RNAi expression plasmids were purchased from Invitrogen. Lenti α‐SMA short hairpin RNA (shRNA), lenti control shRNA, lenti α‐SMA open reading frame vector, lenti PTEN open reading frame vector, and lenti control vector were purchased from the Functional Genomics Facility of University of Colorado. Lipofectamine 2000 was purchased from Life Technologies, Inc. Runx2 3′ untranslated region (UTR) Lenti reporter–Luc vector, the Luciferase Reporter Assay Kit, and TransDux transduction reagent were purchased from System Biosciences. MK‐2206 was purchased from Selleck Chemicals. Medium 199 was purchased from Lonza. Osx 3′ UTR lenti reporter–Luc vector and all other chemicals and reagents were from Sigma‐Aldrich.