Protease inhibitors and pvdf membranes

PRIR-REPORT luciferase reporter vector, miR-124a mimic/control and miR-124a inhibitor/control (Ambion, New York, NY, USA); lymphocyte isolation liquid (Solarbio, New York, NY, USA); Human T Cell Nuclear Transfection kit (Lonza, Bern, Switzerland); MACSRCD4 MicroBeads (Miltenyi Biotec, Bergisch Gladbach, Germany); QuickChange Lighting Mutagenesis kit (Stratagene, La Jolla, CA, USA); TRIzol reagent, PrimeScript^® RT reagent kit with gDNA Eraser and SYBR^® Premix Ex Taq™ II (Takara Biotechnology Co., Ltd., Dalian, China); protease inhibitors and PVDF membranes (Roche Diagnostics, Basel, Switzerland); Dual-Luciferase^® Reporter (DLR™) Assay kit (Promega, Madison, WI, USA); ELISA kit (R&D Systems, Minneapolis, MN, USA); SIRT1, β-actin mouse monoclonal antibody, goat anti-mouse HRP antibody (Cell Signaling Technology, Inc., Boston, MA, USA); modified BCA assay kit (Sangon, Shanghai, China). All the primers were synthesized by Sangon.

Human OSCC cell lines SCC15 and Cal27 were generously presented by the Department of Stomatology, Wuhan University (Wuhan, China). Dulbecco's modified Eagle's medium (DMEM) and fetal bovine serum (FBS) (both from Gibco, Grand Island, NY, USA); TRIzol reagent, PrimeScript^® RT reagent kit with gDNA Eraser and SYBR^® Premix Ex Taq™ II (Takara Biotechnology Co., Ltd., Dalian, China); RIPA lysate (Thermo Fisher Scientific, Inc., Carlsbad, CA, USA); protease inhibitors and PVDF membranes (Roche, Basel, Switzerland); rabbit polyclonal TCF-4 antibody (dilution, 1:500; cat. no. ab185736), rabbit monoclonal cyclin D1 antibody (dilution, 1:500; cat. no. ab16663), rabbit polyclonal GAPDH antiboody (dilution, 1:500; cat. no. ab37168) and secondary goat anti-rabbit (HRP) IgG antibody (dilution, 1:5,000; cat. no. ab6721) were all purchased from Abcam (Cambridge, MA, USA). Cell Counting Kit-8 (CCK-8) (Dojindo, Kumamoto, Japan); Lipofectamine 3000 (Invitrogen Life Technologies, New York, NY, USA); Annexin V-FITC cell apoptosis detection kit (eBioscience, San Diego, CA, USA); 5-aza-2′-deoxycytidine (5-aza-CdR; Sigma, New York, NY, USA); TOP/FOP-Flash luciferase reporter vector (Millipore, Billerica, MA, USA); Dual-Luciferase assay kit (Promega, Madison, WI, USA); primers (Sangon, Shanghai, China).