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. 2013 Oct 7;8(10):e74847. doi: 10.1371/journal.pone.0074847

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© 2013 Kooij et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Samples were taken at different time points and run on a 1D gradient gel stained with ProQ Diamond and with Sypro Ruby (M: Peppermint molecular weight marker). (B) Time course of PKCα-mediated phosphorylation of recombinant cTn (DD) complex. Mean values are shown (±S.E.M.) obtained from two different 1D-gels; the intensities of the cTnI bands on the Sypro Ruby stained gels were used to correct for loading differences. Time constant (±S.E.M.) of the exponential fitted to the data points: 60±3 minutes for cTnT and 58±4 minutes for cTnI phosphorylation. (C) Western blotting of the cTn (DD) complex incubated with PKCα for 90 minutes (cTn (DD+PKCα)) showed phosphorylation at cTnI sites Ser42 and Thr143 with phospho-specific antibodies.