Abt 263

Eμ-myc, Eμ-myc/p53^−/−, Eμ-myc/p19^Arf−/−, Eμ-myc/Apaf1^−/− and Eμ-myc/Bcl-2 B-cell lymphomas were kindly provided by Ricky W Johnstone. Eμ-myc lymphomas were cultured in six-well plates in high-glucose Dulbecco's modified Eagle's medium supplemented with 10% fetal calf serum, penicillin/streptomycin, 0.1 mM l-asparagine and 50 μM 2-mercaptoethanol in a 37 °C, 10% CO2 humidified incubator. All human B-cell lymphoma cell lines (RAJI, RL, U2932, RAJI-4RH, RL-4RH and U2932-4RH) were kindly provided by Francisco J Hernandez-Ilizaliturri and cultured under standard conditions (37 °C, with 5% CO2) in RPMI (Roswell Park Memorial Institute) 1640 medium supplemented with 10% fetal bovine serum (FBS) and penicillin/streptomycin. I-BET762 was purchased from XcessBio (San Diego, CA, USA) as a 10 mM DMSO solution. ABT-263 was purchased from ChemieTek (Indianapolis, IN, USA) and dissolved in DMSO for the preparation of stock solutions (10 mM). Obatoclax was purchased from SelleckChem (Houston, TX, USA) as a 10 mM DMSO solution.

Most of the experiments presented here were performed at least twice. Furthermore, some of the experiments were replicated in different cell lines but with different doses and time points, which were determined for each cell line in preliminary experiments.
The anti-P-PI3K antibody was obtained from Santa Cruz, the polyclonal anti-PI3K was from Pierce, the anti-HIF1a antibody was from BD Transduction Laboratories, the cytochrome c antibody was from BD Pharmingen (Cat. 556433 for blots), and the anti-β-tubulin antibody was from BD Bioscience. For pY-IGF1R, we used the anti-pY1135/1136 IGF1R antibody from CST (Cat#3024), and for pS-AKT, we used the anti-pSer 473 AKT1 antibody from Upstate (Cat#05-736). All other primary antibodies were purchased from Cell Signal (CST). Secondary antibodies conjugated with HRP were purchased from GE Healthcare. Most of these antibodies have been tested for specificity by us and other groups. ABT-263 was purchased from ChemieTek; 2-Deoxy-D-glucose was purchased from Sigma. The pan-caspase inhibitor z-VAD was purchased from Promega. Picropodophyllin was purchased from COSMO Bioscience. PD98509, an ERK inhibitor, was purchased from Merck, and LY29402, a PI3K inhibitor, was purchased from CST.

Recombinant L-asparaginase from Escherichia coli was purchased from Sigma Aldrich (St. Louis, MO, U.S.A.) or from ProSpec-Tany Techno Gene Ltd. (Rehovot, Israel). A 500 IU/ml working solution in PBS was prepared prior to storage at −20°C. ABT263 was purchased from ChemieTek (Indianapolis, IN, U.S.A.). A 10 mM working solution in dimethylsulfoxide (DMSO) was prepared prior to storage at −20°C. Recombinant TRAIL was purchased from Peprotech (Rocky Hill, NJ, U.S.A.). A 100 μg/ml working solution in PBS was prepared prior to storage at −20°C.

ATN-224 was provided by Dr Andrew Mazar (Northwestern University, Evanston, IL). ABT-263 and ABT-737 were purchased from ChemieTek (Indianapolis, IN). ZVAD-FMK was purchased from Enzo Life Sciences (Plymouth Meeting, PA). All other drugs and chemicals were purchased from Sigma Chemical Co. (St. Louis, MO) unless otherwise stated.