Mk 2206

The complete NCI-60 panel was obtained from National Cancer Institute (NCI) Anticancer Drug Screen (Bethesda, MD, USA). Pancreatic cell lines were obtained from ATCC (Manassas, VA, USA). All cells were cultivated in RPMI including 2 mM glutamine and 11.11 mM glucose supplemented by 10% FBS and penicillin/streptomycin (GIBCO, Grand Island, NY, USA).
Romidepsin (depsipeptide, NSC 630176) was provided by the Cancer Therapy Evaluation Program, NCI. Tariquidar was provided by the NCI Anticancer Drug Screen. PD-0325901 (MEK inhibitor), MK-2206 (AKT inhibitor) were purchased from ChemieTek (Indianapolis, IN, USA). Glutaminase inhibitor 968 compound was obtained from Millipore (Burlington, MA, USA); oligomycinA, NAC, glutathione, dimethyl-α-ketoglutarate, dimethyl-glutamate, sodium citrate, and sodium acetate were obtained from Sigma-Aldrich (St. Louis, MO, USA); DPI, ML171, and etomoxir was obtained from Selleckchem (Houston, TX). MitoQ was provided by Dan L. Sackett (NIH, NICHD).
Unless otherwise specified, t-tests were performed to evaluate significance of results, and asterisks were added to graphs when the p-value was lower than 0.05.

Chemical and molecular biology reagents were purchased from Fisher Scientific (Pittsburgh, PA, USA) unless stated otherwise. BMS-754807 was purchased from JiHe Pharmaceutica (Beijing, China) and NBI-31772 from Tocris Bioscience (Ellisville, MO, USA). Liberase TM, Alizarin Red S, ZnCl₂, and batimastat were purchased from Sigma (St. Louis, MO, USA). MK2206 was purchased from ChemieTek (Indianapolis, IN). Rapamycin was purchased from Calbiochem (Gibbstown, NJ). Antibodies (phospho-Akt, phospho-S6, and phospho-Erk) were purchased from Cell Signaling Technology (Danvers, MA, USA). Restriction enzymes were purchased from New England BioLabs (Ipswich, MA, USA). Primers, cell culture media, antibiotics, TRIzol, M-MLV reverse transcriptase, Alexa Fluor 488 Tyramide SuperBoost Kit, and Tetramethylrhodamine (TMRM) were purchased from Life Technologies (Carlsbad, CA, USA). Anti-Digoxigenin-POD and Anti-Digoxigenin-AP antibodies were purchased from Roche (Basel, Switzerland).

All chemicals and reagents were purchased from Fisher Scientific (Pittsburgh, PA) unless otherwise specified. NVP-AEW541 was obtained from Novartis (Basel, Switzerland) and BMS-754807 was purchased from JiHe Pharmaceutica (Beijing, China). Wortmannin and LY294002 were purchased from Cell Signaling Technology (Danvers, MA). AZD8055 and Torin1 were purchased from Selleck Chemicals (Houston, TX). Rapamycin and Akti-1/2 were purchased from Calbiochem (Gibbstown, NJ). MK2206 was purchased from ChemieTek (Indianapolis, IN). All inhibitors were dissolved in DMSO and further diluted in water. Restriction enzymes were purchased from New England Bio Labs (Beverly, MA). The TRIzol reagent, M-MLV reverse transcriptase, and RNaseOUT™ Recombinant Ribonuclease Inhibitor were purchased from Invitrogen (Waltham, MA). The anti-GFP antibody was purchased from Torrey Pines Biolabs (Secaucus, NJ). The Phospho-Akt (Ser 473) and Phospho-S6 antibody were purchased from Cell Signaling Technology (Beverly, MA).

Romidepsin was obtained from the Developmental Therapeutics Program of the National Cancer Institute (Bethesda MD). The MEK inhibitor PD-0325901, the AKT inhibitor MK-2206, the ERK inhibitor Vx-11e and the PI3K inhibitor GDC-0941 were purchased from ChemieTek (Indianapolis, IN). Q-VD-OPh was from R&D Systems (Minneapolis, MN). The dual pathway inhibitor, D-87503, was synthesized in-house by the Chemical Biology Laboratory, Center for Cancer Research (Frederick, MD).