FIG 5.
TgMAPK2 degradation impairs centrosome duplication after basal migration. (A) Z-projections of confocal stacks of TgMAPK2AID parasites stably expressing mVenus-TgCentrin1 (green) and TgCEP250L1-3×HA (red) grown for 8 h in the presence (+) or absence (−) of 500 μM IAA and costained with Hoechst stain (blue). Arrows, TgCentrin1; large arrowheads, TgCEP250L1. Nuclei are outlined in yellow. (B) Quantification of parasites with duplicated centrosomal inner cores (TgCEP250L1) (B) and outer cores (TgCentrin1) (C) grown in the presence or absence of IAA for 8 h. Means ± SD from 3 biological replicates are shown; 200 to 500 parasites were counted under each condition. (D) Maximum-intensity Z-projections of confocal stacks of TgMAPK2AID parasites stably expressing mVenus-TgCentrin1 (green) and GRASP55-mRuby3 (red) during the cell cycle. Representative images summarizing centrosome migration/duplication are shown. (E) Quantification of parasites in each category shown in panel D grown in the presence or absence of IAA for 8 h. Means ± SD from 3 biological replicates are shown; 300 to 500 parasites were counted under each condition. P values are from two-tailed Student’s t test. Bars = 2 μm.