LS174T, MDA MB 231 cells, RPMI and DMEM media were from Clare Hall labs, UK. Fetal bovine serum Gold was from PAA Laboratories. Tet Program Accepted FBS was from Docetaxel clinical trial ClontechTakara Bio Europe, St Germain en Laye, France. Protein Aagarose was from Santa Cruz Biotechnology Inc. Heidelberg, Germany. OligofectamineTM and OptimemTM were from Invitrogen Ltd. Paisley, UK. Lambda Protein Phosphatase was from New England BioLabs, Hitchin, UK. Protease inhibitor cocktail tablets were from Roche Diagnostics Ltd. Burgess Hill, UK. RIPA load, Triton X100 and phosphatase inhibitor cocktail 1 and 2 were from SigmaAldrich Company Ltd. Gillingham, UK. DRAQ5 was from Alexis Biochemicals and Immobilon P PVDF membrane was from Millipore. The PhosphoProtein Refinement system was from Qiagen Ltd. Crawley, UK. Fluoromount was from Dako, Ely, UK. Akt, Phospho Akt Phospho Bcl 2 PhosphoBcl Infectious causes of cancer 2 Bcl xL, JNK, Phospho JNK PARP, Phospho vimentin p70 S6 kinase, phospho p70 S6 kinase were from Cell Signalling Technology. BNIP3 and BNIP3L were from SigmaAldrich Ltd. Gillingham, UK. Cyclin B1 and HIF 1a were from BD Pharmingen, Oxford, UK. Bcl 2, Bax and Tom20 were from Santa Cruz Biotechnology Inc. Heidelberg, Germany and TrueBlotTM Ultra HRP anti mouse IgG was from eBioscience Ltd. Hatfield, UK. Alexa Fluor 488 goat anti mouse IgG2b and Alexa Fluor 546 goatantimouse IgG2a were from Invitrogen Ltd. Paisley, UK. Doxorubicin hydrochloride, vinblastine sulfate, paclitaxel, doxycycline hyclate, vinorelbine ditartrate salt, colchicine, nocodazole and sulforhodamine W sodium salt were from SigmaAldrich Ltd. Gillingham, UK. Rapamycin and SP600125 were from Calbiochem. Gossypol molecular weight Bortezomibwas something special from Millennium Pharmaceuticals, Cambridge, MA, USA. Cisplatin was a gift from Dr. Richard Callaghan, University of Oxford. BNIP3 RNAi duplexes were designed using Dharmacon siDESIGN1 centre scrambled get a grip on RNAi duplexes were designed using InvivoGen siRNA wizardTM and all RNAi duplexes were synthesised and annealed by Eurogentec, Seraing, Belgium. Tetracycline inducible HCT116 cells were prepared utilizing the flp in T Rex process based on the manufacturers directions. Hypoxic incubations were conducted having an INVIVO2400hypoxicworkstation. To create a combined anoxic/low pH atmosphere, cells were placed in an jar containing an sachet at 37 8C for the duration of the test. Cells were washed with 4 8C PBS and homogenised on ice in lysis buffer containing 6. 2 Murea, 10% glycerol, 5 mMDTT, 1000 SDS, protease inhibitors and phosphatase inhibitors. Proteins were separated by SDS PAGE, using 12% TrisHCl ties in.