FIGURE 4.
Plasmodium homologue of the Mini-Chromosome Maintenance Complex Binding Protein (PfMCMBP)-deficient parasites complete budding and form merozoites with varied cell body size and ploidy. (A) Representative pictures from [+]/[−] Shld1 PfMCMBP3HADDparasites treated with E64, stained with SYBR Green I and probed with rabbit anti-PfGAP45. Images were taken with LSM880 Airyscan using the 60× objective and the scale bar is at 1 μm. (B) The measurement of cytoplasm and nucleus area were obtained using Image J software. Three biological replicates with a total of 284 and 392 merozoites were, respectively, measured for [+] and [−] Shld1 conditions. (C) The ploidy per nucleus was obtained using Image J software and calculating the corrected total cell fluorescence (CTCF) of the SYBR Green I fluorescence intensity calculated using the following the formula: CTCF = Integrated Density - (Area of selected cell × Mean fluorescence of background readings). Three biological replicates and a total of 306 and 361 merozoites were respectively counted for [+] and [−] Shld1 conditions. (D) Representative still images from time lapse microscopy of [+]/[−] Shld1 PfMCMBP3HADDparasites during egress. Scale bar at 5 μm