Fig. 1: PfAnchor is an essential conserved Plasmodium protein expressed through the asexual life cycle.

a Schematic representation of PfAnchor’s structural features and predicted protein folding. PfAnchor primary sequence shows a single phosphorylation site at S204. Alphafold3 modeling combined with a DALI protein domain search identified a conserved pleckstrin homology (PH) domain (PfAnchor AA 768-998) across Plasmodium species. The sequence alignment of this region shows strong conservation, with a predicted phosphoinositides (PIP)-binding motif (PfAnchor AA 892-904) highlighted in magenta in the structural model. b Schematic of PfAnchor tagging strategy and conditional knockdown system. PfAnchor was tagged with an smV5 epitope for visualization, and knockdown was achieved using the TetR-aptamer system, where protein translation depends on anhydrotetracycline (aTC) availability. c Representative western blot from one of three independent experiments performed on separate parasite protein lysates from ring (R), trophozoite (T), and schizont (S) stage. Anti-V5 detects PfAnchor, showing that it is expressed throughout the asexual life cycle with highest abundance in schizont stages. Anti-PfAldolase is used as a loading control, as it is a constitutively expressed cytoplasmic protein that maintains relatively stable levels across all intraerythrocytic stages. d Representative western blot from one of four independent experiments performed on separate parasite cultures confirming PfAnchor knockdown efficiency. Quantification performed via measurement of the fluorescent intensities of the sample (V5) bands compared to the normalized intensities of the loading control (PfAldolase) bands. Plotted is mean +/- SD, n = four biological replicates. Molecular weight markers are indicated on the left. Cropped regions are from the same blot. Full, uncropped scans are provided in the Supplementary Fig. 17. Source data are provided as a Source Data file.