%0 Journal Article %A Brahimi-Horn, M. %A Giuliano, Sandy %A Saland, Estelle %A Lacas-Gervais, Sandra %A Sheiko, Tatiana %A Pelletier, Joffrey %A Bourget, Isabelle %A Bost, Frédéric %A Féral, Chloé %A Boulter, Etienne %A Tauc, Michel %A Ivan, Mircea %A Garmy-Susini, Barbara %A Popa, Alexandra %A Mari, Bernard %A Sarry, Jean-Emmanuel %A Craigen, William %A Pouysségur, Jacques %A Mazure, Nathalie %D 2015 %T Additional file 18: Figure S10. of Knockout of Vdac1 activates hypoxia-inducible factor through reactive oxygen species generation and induces tumor growth by promoting metabolic reprogramming and inflammation %U https://springernature.figshare.com/articles/journal_contribution/Additional_file_18_Figure_S10_of_Knockout_of_Vdac1_activates_hypoxia-inducible_factor_through_reactive_oxygen_species_generation_and_induces_tumor_growth_by_promoting_metabolic_reprogramming_and_inflammation/4475129 %R 10.6084/m9.figshare.c.3646397_D3.v1 %2 https://ndownloader.figshare.com/files/7205030 %K Wt RAS MEF %K Soft agar assay %K RASV %K FCCP %K OCR %K Glu 10  mM %K reactive oxygen species generation %K oxygen consumption rate %K ECAR %K Oligo 1 μ M %K Nx %K 1 μ %K X μ m %K extracellular acidification rate %K WT RAS MEF %K Representative phase contrast photographs %K clonogenic cell survival assay %K Seahorse XF bioenergetic system %K Vdac 1 activates hypoxia-inducible factor %X Characterization of RASV12 transformed MEF expressing (WT RAS MEF) or not (Vdac1 −/− RAS MEF) Vdac1. (A) Characterization of the growth of Wt and Vdac1 −/− RAS MEF incubated in Nx or Hx for the indicated number of days. The mean ± SEM is representative of four independent experiments carried out in duplicate. A p < 0.00001 shows significant difference from the normoxia for Vdac1 −/− RAS MEF. (B) Representative phase contrast photographs of Wt and Vdac1 −/− RAS MEF incubated in Nx for 72 h. Scale bars represent X μm. (C) Relative migration of Wt and Vdac1 −/− RAS MEF in Nx as evaluated in a xCELLigence system. The mean ± SEM is representative of two independent experiments carried out in quadruplicate. (D) The extracellular acidification rate (ECAR) in Nx of Wt and Vdac1 −/− RAS MEF was evaluated with a Seahorse XF bioenergetic system. Glucose (Glu 10 mM) and oligomycin (Oligo 1 μM) were injected at the indicated times. (E) The oxygen consumption rate (OCR) in Nx for Wt and Vdac1 −/− RAS MEF was measured in real time with a Seahorse XF. Glucose (Glu 10 mM), oligomycin (Oligo 1 μM), carbonilcyanide p-triflouromethoxyphenylhydrazone (FCCP 1 μM), and Rotenone/Antimycine A (Rot/AA, 1 μM/1 μM) were injected at the indicated times. The mean ± SEM is representative of three independent experiments carried out in quadruplicate. (F) Radioresistance of Wt and Vdac1 −/− RAS MEF cultured for 24 h in Nx or Hx and treated with the indicated dose of radiation. Cell growth was then evaluated with a clonogenic cell survival assay. X-axis: dose of X-radiation (Gy). Y-axis: surviving fraction. The mean ± SEM is representative of two independent experiments carried out in duplicate. (G) Soft agar assay of Wt and Vdac1 −/− RAS MEF. (H) Tumor weight of Wt (Wt RAS MEF) and Vdac1 −/− RAS MEF-derived tumors (Vdac1 −/− RAS MEF). %I figshare