Sevoflurane promotes the expansion of glioma stem cells through activation of hypoxia- inducible factors in vitro Q.Y. Shi, S.J. Zhang, L. Liu, Q.S. Chen,

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Sevoflurane promotes the expansion of glioma stem cells through activation of hypoxia- inducible factors in vitro Q.Y. Shi, S.J. Zhang, L. Liu, Q.S. Chen, L.N. Yu, F.J. Zhang, M. Yan British Journal of Anaesthesia Volume 114, Issue 5, Pages 825-830 (May 2015) DOI: 10.1093/bja/aeu402 Copyright © 2015 The Author(s) Terms and Conditions

Fig 1 GSCs exposed to either 2% sevoflurane or NC. (a) Representative image of EdU staining (red) with Hoechst 33342 counterstain (blue). (b) Representative image of sphere-forming ability and number of spheres (mean and sd, **P<0.01). British Journal of Anaesthesia 2015 114, 825-830DOI: (10.1093/bja/aeu402) Copyright © 2015 The Author(s) Terms and Conditions

Fig 2 Representative images of western blots of CD133, VEGF, HIFs, p-Akt, and β actin after (a) exposure of GSCs to 2% sevoflurane for up to 6 h. (b) Exposure to 0–6% sevoflurane for 4 h. HIF, hypoxia induced factor; NC, naïve control; Sevo, sevoflurane. British Journal of Anaesthesia 2015 114, 825-830DOI: (10.1093/bja/aeu402) Copyright © 2015 The Author(s) Terms and Conditions

Fig 3 Representative western blots of HIFs and β actin expression and percentage of EdU-positive cells after exposure of GSCs to 2% sevoflurane for 6 h (mean and sd, **P<0.01, #P<0.05) after treatment with (a) HIF-1α siRNA and (b) HIF-2α. Cont, control siRNA. British Journal of Anaesthesia 2015 114, 825-830DOI: (10.1093/bja/aeu402) Copyright © 2015 The Author(s) Terms and Conditions

Fig 4 Effects of exposure of GSCs to 2% sevoflurane with and without the Akt inhibitor, LY294002. (a) Representative western blots of CD133, HIFs, Akt, and β actin expression, (b) representative image of EdU staining (red) and Hoechst 33342 counterstain (blue) and (c) percentage of EdU-positive cells (mean and sd, **P<0.01, #P<0.05). NC, naïve control; Sevo, sevoflurane; LY, LY294002. British Journal of Anaesthesia 2015 114, 825-830DOI: (10.1093/bja/aeu402) Copyright © 2015 The Author(s) Terms and Conditions