Figure 6: Phosphorylation of PfCDPK1 at S64 occurs during the schizont (Schiz) stage of parasite development. | Nature Communications

Figure 6: Phosphorylation of PfCDPK1 at S64 occurs during the schizont (Schiz) stage of parasite development.

From: Phosphoproteomics reveals malaria parasite Protein Kinase G as a signalling hub regulating egress and invasion

Figure 6

(a) Parasite lysates (15 μg) prepared from ring, trophozoite (Troph) or Schiz stage and were probed with the structural PfCDPK1 antibody, the phospho-specific CDPK1-pS64 antibody or the Schiz-specific marker PfEBA-175. (b) A late Schiz culture treated with vehicle or Compound 2 (2μM; 60min) 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 phospho-specific 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 images (see; Supplementary Movie 2—video for z stacks) are of a free merozoite. These results are representative of at least three experiments. Scale bars, b=1 μm, c=0.5 μm, d=0.5 μm, e=1 μm, f=0.5 μm

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