Mouse monoclonal anti cyclin b1

Cell lysis and Western blot analysis were performed as described previously [40 (link)], using the following antibodies: Rabbit polyclonal anti-SIK2 (Cell Signaling, Danvers, MA, USA), rabbit polyclonal anti-SIK2-pS385 (Kinexus, Sydney, Australia), rabbit polyclonal anti-AKT (Cell Signaling), rabbit polyclonal anti-AKT-pS473 (Cell Signaling), mouse monoclonal anti-β-actin (Sigma-Aldrich), rabbit polyclonal-anti KIF18B (ThermoFisher scientific, Waltham, MA, USA), rabbit polyclonal anti-EB1 (Abcam, Cambridge, UK), mouse monoclonal anti-cyclin B1 (Santa Cruz Biotechnology, Dallas, TX, USA), mouse monoclonal anti-Histone 3-pS10 (ThermoFisher scientific), mouse monoclonal anti-PLK1 (Santa Cruz Biotechnology), mouse monoclonal anti-vinculin (Santa Cruz Biotech), mouse monoclonal anti-P53 (Santa Cruz Biotech), rabbit polyclonal anti-P21 (cell signaling), rabbit polyclonal anti-BCL-xL (Santa Cruz Biotechnology), rabbit polyclonal anti-Cyclin D (Santa Cruz Biotechnology).
Reagents were purchased from the following sources: Paclitaxel (T7402) Sigma-Aldrich, propidium iodide (440300250) Acros Organics (Fair Lawn, NJ, USA), RNase A (1007885) Qiagen (Hilden, Germany), PE Annexin V (556421) and 7AAD (21-68981E) BD Biosciences, Caspase-Glo^® 3/7 Assay system (P1781) Promega, RO3306 (S7747) Selleckchem (Houston, TX, USA).

Western blot analysis was performed as described [47 (link)-49 (link)] and the following primary antibodies were used: mouse monoclonal anti-Kif2c (Santa Cruz biotechnology, Heidelberg), mouse monoclonal anti-Plk1 (Santa Cruz biotechnology), mouse monoclonal anti-cyclin B1 (Santa Cruz biotechnology), mouse monoclonal anti-β-actin (Sigma-Aldrich) and mouse monoclonal anti-Hec1 (Abcam, Darmstadt). Cell cycle was analyzed as described [17 (link),19 (link)].

Sodium dichloroacetate (DCA), metformin hydrochloride, 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), crystal violet, RNAse, propidium iodide (PI), RIPA buffer and protease inhibitor cocktails were obtained from Sigma (St. Louis, MO, USA). Mouse monoclonal anti-cyclin D3 antibody was purchased from Cell Signaling Technology, Inc. (Beverly, MA, USA). Goat anti-rabbit IgG-HRP, goat anti-mouse IgG-HRP, mouse monoclonal anti-cyclin B1 and anti-cytochrome C antibodies were obtained from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Monoclonal anti β-actin antibody was obtained from Sigma (St. Louis, MO, USA). Mouse monoclonal anti-PDH-E1α was purchased from Abcam (Cambridge MA, USA). Rabbit polyclonal PhosphoDetect^TM anti-PDH-E1α (pSer³⁰⁰) was purchased from Calbiochem (EMD Millipore Corp., MA, USA).