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. 2013 May 31;452(Pt 3):457–466. doi: 10.1042/BJ20121694

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© 2013 The author(s) has paid for this article to be freely available 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 License, which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited.

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(A) Western blot analysis of transgenic CK2–HA parasites that express an endogenous (3D7–CK2–HA) fusion protein. Expression of the fusion proteins was confirmed using an anti-HA antibody. The parental 3D7 strain (3D7) was used as a control. (B) Immunofluorescence microscopy using endogenously derived PfCK2α–HA. Localization with the anti-HA antibodies (green) confirmed expression in the nucleus (blue, DAPI) and cytosol of fixed parasites in the trophozoite (T) and schizont (S) stage. Scale bar indicates 2 μm. Enlargements of selected areas are marked with a white square. DIC, differential interference contrast microscopy. (C) Western blot analysis of transgenic CK2–HA parasites that overexpress CK2–HA. Expression of the fusion proteins was confirmed using an anti-HA antibody. This cell line was used to immunopurify CK2–HA for subsequent phosphorylation assays. (D) Western blot analysis of phosphorylated Rh2b CPD using immunoprecipitated CK2–HA. Phosphorylation was detected using the phospho-specific anti-pRh2b antibody. CK2 was visualized with an anti-HA antibody and an anti-GST antibody was used as the loading control. (E) Western blot analysis of CK2-dependent phosphorylation of Rh2b in the presence of different heparin concentrations. CK2 was visualized with an anti-HA antibody and an anti-GST antibody was used as the loading control. (F) Western blot analysis of CK2-dependent phosphorylation of Rh2b CPD in the presence of different TBCA concentrations. CK2 was visualized with an anti-HA antibody and an anti-GST antibody was used as the loading control. The molecular mass is shown in kDa on the left-hand side of the blots.