P38alpha kinase plays an important role in the regulation of both cell stress response and cell fate. In this study, we report that p38alpha kinase-deficient embryonic stem cells exhibit a higher production of reactive oxygen species (ROS) in contrast to their wild-type counterpart. Analysis of the expressions of NADPH oxidases (NOXs) and dual oxidases, crucial enzymes involved in intracellular ROS formation, shows NOX2/gp91phox is over-expressed in p38alpha deficient cells. The particular increase in superoxide formation was confirmed by the specific detection of hydroethidine derivate 2-hydroxyethidium. ROS formation decreased when the level of NOX2 was silenced by siRNA in p38alpha deficient cells. These data suggest the importance of p38alpha kinase in the regulation of ROS metabolism in embryonic stem cells and the significance of the observed phenomena of cancer cell-like phenotypes, which is discussed.

Department of Cytokinetics Institute of Biophysics Academy of Sciences of the Czech Republic Brno Czech Republic

Department of Free Radical Pathophysiology Institute of Biophysics Academy of Sciences of the Czech Republic Brno Czech Republic

Institute of Experimental Biology Faculty of Science Masaryk University Brno Czech Republic

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$a Binó, Lucia $u Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic; Department of Free Radical Pathophysiology, Institute of Biophysics, Academy of Sciences of the Czech Republic, Brno, Czech Republic. Electronic address: bino@ibp.cz.

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$a The depletion of p38alpha kinase upregulates NADPH oxidase 2/NOX2/gp91 expression and the production of superoxide in mouse embryonic stem cells / $c L. Binó, I. Veselá, I. Papežíková, J. Procházková, O. Vašíček, K. Štefková, J. Kučera, M. Hanáčková, L. Kubala, J. Pacherník,

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$a P38alpha kinase plays an important role in the regulation of both cell stress response and cell fate. In this study, we report that p38alpha kinase-deficient embryonic stem cells exhibit a higher production of reactive oxygen species (ROS) in contrast to their wild-type counterpart. Analysis of the expressions of NADPH oxidases (NOXs) and dual oxidases, crucial enzymes involved in intracellular ROS formation, shows NOX2/gp91phox is over-expressed in p38alpha deficient cells. The particular increase in superoxide formation was confirmed by the specific detection of hydroethidine derivate 2-hydroxyethidium. ROS formation decreased when the level of NOX2 was silenced by siRNA in p38alpha deficient cells. These data suggest the importance of p38alpha kinase in the regulation of ROS metabolism in embryonic stem cells and the significance of the observed phenomena of cancer cell-like phenotypes, which is discussed.

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$a Veselá, Iva $u Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic. Electronic address: vesna@mail.muni.cz.

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$a Papežíková, Iva $u Department of Free Radical Pathophysiology, Institute of Biophysics, Academy of Sciences of the Czech Republic, Brno, Czech Republic. Electronic address: papezikova@ibp.cz.

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$a Procházková, Jiřina $u Department of Cytokinetics, Institute of Biophysics, Academy of Sciences of the Czech Republic, Brno, Czech Republic. Electronic address: iris@ibp.cz.

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$a Vašíček, Ondřej $u Department of Free Radical Pathophysiology, Institute of Biophysics, Academy of Sciences of the Czech Republic, Brno, Czech Republic. Electronic address: ondrej.vasicek@ibp.cz.

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$a Štefková, Kateřina $u Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic. Electronic address: 150806@mail.muni.cz.

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$a Kučera, Jan $u Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic. Electronic address: 150663@mail.muni.cz.

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$a Hanáčková, Markéta $u Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic. Electronic address: 17059@mail.muni.cz.

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$a Kubala, Lukáš $u Department of Free Radical Pathophysiology, Institute of Biophysics, Academy of Sciences of the Czech Republic, Brno, Czech Republic. Electronic address: kubalal@ibp.cz.

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$a Pacherník, Jiří $u Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic. Electronic address: jipa@sci.muni.cz.

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