Cells were then rinsed with PBS and grown for 7–10 days in complete DMEM supplemented with feeder cells (V79 cells exposed to 250 Gy 60Co irradiation). Construction details are given in Materials and methods. The backbone vector pCI-puro was made by replacing the neomycin resistance cassette from pCI-neo (Clontech, Basingstoke, UK) with the puromycin resistance gene from pPUR (Clontech) as a PvuII/BamHI fragment (all enzymes from Life Technologies). Hypoxia induces c-fos transcription via a mitogen-activated protein kinase-dependent pathway J Biol Chem 1997 272: 23435–23439, Yan SF et al. T24 cells were transiently transfected with these constructs and their sensitivity to the prodrug indole-3-acetic acid (IAA) was examined. analyzed data; and R.O.-C., J.C.-G., and J.L.-B. Oxidative stress produced by suprahepatic occlusion and reperfusion Hepatology 1993 18: 881–889, Chandel NS et al. Surviving fractions were normalized for plating efficiency of cells exposed to HBSS only. We are very grateful to Dr Gill Tozer, Professor Mike Joiner and Professor Ian Stratford for helpful discussion, and to Mrs Claudia Coralli and Mrs Sara Bourne for excellent technical support. Implications for the mechanism of oxygen sensing Eur J Biochem 1995 234: 92–99, Huang LE, Arany Z, Livingston DM, Bunn HF . In contrast, the full-length CMV promoter was not affected by the treatments in MCF-7 cells, while slightly down-regulated in T24 cells Figures 2Figures 3Figures 4Figures 5. NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. The addition of a further five copies of the PGK1 HRE in the construct pPGK110/E9 did not significantly increase the hypoxia-inducibility in T24 cells (fold-induction 1.7 ± 0.3). IAA was purchased from Sigma. Gene therapy approaches based on the selective expression of therapeutic genes present a number of distinctive advantages.1718 Artificial hybrid promoters can offer specificity, as well as applicability in a wide range of cancer types, complementing and enhancing the properties of the different components.19 They are also characterized by flexibility, because several elements of different origin can be arranged in various orders, numbers and relative orientations, in order to optimize gene regulation. Astronomers ostensibly know plenty about neutron stars. PubMed Google Scholar. Finally, efficient and selective prodrug-mediated toxicity was observed when the Epo/E9 promoter controlled the expression of HRP after combined hypoxia and radiation treatment Figure 5. Conversely, after hypoxic and/or radiation treatment, selective toxicity was only seen when the stimuli-inducible promoters were employed. Localization of the protein product of the immediate early growth response gene, Egr-1, in the kidney after ischemia and reperfusion Cell Regul 1991 2: 251–260, Shibata T, Giaccia AJ, Brown JM .