1. In vivo phosphorylation of S3233 (S3233 is the sole phosphoacceptor site in the cytoplasmic domain of Rh2B for in vitro phosphorylation by parasite extract) in schizonts . Expression was verified by IFA using anti-HA antibodies. Scale bar indicate 2 μm. Rh2B is localized at the apical pole of nascent merozoites.

2. Expression, purification of PfCK2 alpha (endogenously HA-tagged (3D7-CK2-HA, PF3D7_1108400) and Rh2b in vitro phosphorylation. Immuno-fluorescence microscopy using endogenous derived PfCK2-HA alpha. 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 the endogenous tagged protein was localized in the parasite´s cytosol, as well as in the nucleus of trophozoite stage parasites. In the schizont stage of the parasite the dual localization of endogenous tagged CK2 was less pronounced and appeared predominantly cytosolic.

Engelberg K, Paul AS, Prinz B, Kono M, Ching W, Heincke D, Dobner T, Spielmann T, Duraisingh M, Gilberger TW. Specific phosphorylation of the PfRh2b invasion ligand of Plasmodium falciparum. Biochem J. 2013 452(3):457-66