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. 2014 Aug 6;34(4):e00129. doi: 10.1042/BSR20140079

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© 2014 The Author(s) This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (CC-BY) (http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (CC-BY) (http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited.

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(A) Nitrate/nitrite accumulation in HEK-293 cell culture medium 48 h after plating. The nitrate/nitrite accumulation in culture medium was determined using a colorimetric assay kit obtained from Cayman Chemical Co. (B) L-citrulline formation activity was determined in the cell lysates of various eNOS–HEK-293 stable clones. Reactions were carried out as described in the ‘Materials and Methods’ section. (C) The eNOS–HEK-293 cell lysates were immunoblotted with an antibody specific to total eNOS to confirm the equal expression levels of each eNOS construct. Data shown are the mean±S.D. (*P<0.05; **P<0.01; NS, no significant difference versus WT eNOS). Each experiment was performed in triplicate and repeated three times.