Immunoblotting Assays: Antibody Characterization

The following antibodies were used for immunoblotting assays: anti-AKT (#9272), anti-ERK1/2 (#4695), anti-GSK-3B (#9315), anti-MCL-1 (#4572), anti-SYK (#2172) and anti-LYN (#4576) were purchased from Cell Signaling (Danvers, MA, USA). Anti-XIAP (#610762) was purchased from BD Bioscience (San Jose, CA, USA). Anti-HSP90B (#837159A) and anti-GAPDH (#TAB1001) were purchased from Thermo Fisher Scientific (San Jose, CA, USA). Anti-BCL-2 (#k2206) was purchased from Santa Cruz Biotechno-logy (Santa Cruz, CA, USA). Annexin V-fluorescein isothiocyanate and 7-Aminoactinomycin D were obtained from BD Bioscience. The propidium iodide and InnoCyte Flow Cytometric Cytochrome c Release kit were purchased from EMD Millipore (Kankakee, IL, USA). PU-H71 and PU-H71-conjugated agarose beads were obtained from Dr Gabriela Chiosis and were prepared as previously reported.^{17 (link)} Amaxa Solution V Nucleofector kit was purchased from Amaxa (Cologne, Germany). Pools of siRNA against human BTK, AKT and HSP90 were customarily designed and purchased from Thermo Scientific (Waltham, MA, USA).