Obal D. , Weber N. C. , Zacharowski K. , Toma O. , Dettwiler S

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Role of protein kinase C-ε (PKCε) in isoflurane-induced cardioprotection Obal D. , Weber N.C. , Zacharowski K. , Toma O. , Dettwiler S. , Wolter J.I. , Kratz M. , Mu¨llenheim J. , Preckel B. , Schlack W. British Journal of Anaesthesia Volume 94, Issue 2, Pages 166-173 (February 2005) DOI: 10.1093/bja/aei022 Copyright © 2005 British Journal of Anaesthesia Terms and Conditions

Fig 1 Experimental protocol: CON, control group; S, staurosporine 10 μg kg−1. The arrow indicates the time of staurosporine administration; isoflurane was given continuously for 15 min with (0.4MAC+S, 1.0MAC+S, 1.75MAC+S) or without (0.4MAC, 1.0MAC, 1.75MAC) staurosporine. British Journal of Anaesthesia 2005 94, 166-173DOI: (10.1093/bja/aei022) Copyright © 2005 British Journal of Anaesthesia Terms and Conditions

Fig 2 Infarct size (percentage of area at risk) of controls (CON) and isoflurane-preconditioned hearts (0.4MAC, 1MAC, 1.75MAC): S, staurosporine 10 μg kg−1. Data are mean (SEM); n=10 in each group. Infarct size was significantly reduced after isoflurane preconditioning at all concentrations used (†P<0.05 vs CON and S). Attenuation of protection by staurosporine was only observed after administration of 0.4 MAC isoflurane. British Journal of Anaesthesia 2005 94, 166-173DOI: (10.1093/bja/aei022) Copyright © 2005 British Journal of Anaesthesia Terms and Conditions

Fig 3 (a) Representative western blot experiment on cytosolic fraction of controls (CON), isoflurane-treated hearts (0.4MAC, 1MAC, 1.75MAC; n=6 in each group) and groups receiving staurosporine10 μg kg−1 (S) before the preconditioning protocol (0.4MAC+S, 1MAC+S, 1.75MAC+S). Phosphorylated PKCε, total PKCε (i.e. phospho-PKCε and non-phosphorylated PKCε in the cytosolic fraction) and the internal marker α-tubulin are shown. (b) Densitometric evaluation of six experiments as the x-fold increase in average light intensity (AVI) vs control measurements (CON). Data (mean [SEM]) show the ratio of phosphorylated to total PKCε. Only the lowest concentration of isoflurane significantly increased phosphorylation of PKCε (*P<0.05 vs CON), an effect that was abolished by administration of staurosporine. Preconditioning with 1 or 1.75 MAC isoflurane had no effect on PKCε phosphorylation. British Journal of Anaesthesia 2005 94, 166-173DOI: (10.1093/bja/aei022) Copyright © 2005 British Journal of Anaesthesia Terms and Conditions

Fig 4 (a) Cytosolic and (b) membrane fraction of PKCε in controls (CON) and isoflurane preconditioned hearts with (0.4MAC+S, 1.75MAC+S) or without administration of staurosporine (0.4MAC, 1.75MAC), respectively. The data present densitometric evaluation of six experiments as the x-fold increase in average light intensity (AVI) vs control measurement. Only 0.4 MAC isoflurane leads to translocation of PKCε to the membrane fraction, and this effect was blocked by staurosporine. Higher concentrations of isoflurane had no effect on the translocation of PKCε. (*P<0.05 vs CON.) British Journal of Anaesthesia 2005 94, 166-173DOI: (10.1093/bja/aei022) Copyright © 2005 British Journal of Anaesthesia Terms and Conditions