Phosphorylation of PfCDPK1 at S64 occurs during the schizont (Schiz) stage of parasite development. (b) A late Schiz culture treated with vehicle or Compound 2 (2 mM; 60 min) was fixed and probed with DAPI stain to reveal the nuclei (blue), the anti-HA antibody to reveal PfCDPK1-HA expressed in transgenic parasites (red) or the phosphospecific CDPK1-pS64 antibody (green). A merge of images from all three stains is shown as well as a differential interference contrast (DIC) image. (c,d) Free merozoites were fixed and probed with DAPI stain to reveal the nuclei (blue), the anti-HA antibody to reveal PfCDPK1-HA (red) or the phospho-specific CDPK1-pS64 antibody (green). A merge of images from all three stains is shown. (e) A rendered image where deconvoluted z stacks (see; Supplementary Movie 1) were reconstructed in 3D, with interpolation, of wild-type parasites stained with DAPI (blue), the structural CDPK1 antibody to reveal the total pool of PfCDPK1 (red) and the CDPK1-pS64 antibody (green). (f) Same as e but in this case the are of a free merozoite. These results are representative of at least three experiments. Scale bars, b=1 mm, c=0.5 mm, d=0.5 mm, e=1 mm, f=0.5 mm

Alam MM, Solyakov L, Bottrill AR, Flueck C, Siddiqui FA, Singh S, Mistry S, Viskaduraki M, Lee K, Hopp CS, Chitnis CE, Doerig C, Moon RW, Green JL, Holder AA, Baker DA, Tobin AB. Phosphoproteomics reveals malaria parasite Protein Kinase G as a signalling hub regulating egress and invasion. Nat Commun. 2015 6:7285.