FIG 6.

EhCoactosin S147A expression increases the range of E. histolytica movement within a model extracellular matrix. Trophozoites trapped within Matrigel matrix were imaged for 2 h at 2-min intervals, and movement of individual trophozoites was tracked using the manual tracking plugin of NIH ImageJ. Six independent experiments were conducted with the empty vector-, wild type (Coac-WT)-, EhCoactosin S147A (Coac-A)-, and EhCoactosin S147D (Coac-D)-expressing trophozoites, with 18 trophozoites imaged/cell line/experiment. (A) Superimposed motility tracks for the vector control and EhCoactosin WT-, S147A (Coac-A)-, and S147D (Coac-D)-overexpressing trophozoites. Tracks from one experiment are shown (n = 18 per cell line). Pixel coordinates for each trophozoite from the original images were corrected to the midpoints of the extreme x and y axis values for display on a single graph. Pixel length = 0.32 μM. (B) Effect of EhCoactosin S147A and S147D expression on amoebic movement. The distance between the two farthest points was determined for each trophozoite and used to determine the mean range of movement (i.e., maximum distance moved) for each cell line during a given experiment. Each data point represents the mean maximum distance moved for the 18 trophozoites of that cell line measured in one experiment, with data for six biological replicates shown. *, P < 0.03 by one-way ANOVA with Tukey’s multiple-comparison test.