FIG 4 .
Disruption of ROP54 in type II parasites does not affect growth in vitro. (A) IFA results, demonstrating the loss of ROP54HAII staining in a ∆rop54II clone. (B) IFA results for ∆rop54II parasites complemented with ROP54HAII at the ku80 locus (designated ROP54cII). Proper localization of ROP54 in the ROP54cII parasite clone was assessed by colocalization with ROP13. (C) Western blot assay results, demonstrating loss of HA signal in ∆rop54II parasites and restoration of HA signal for ROP54cII parasites. ROP9 is shown as a loading control. (D and E) HFF monolayers were infected with ROP54HAII, ∆rop54, or ROP54cII parasites, and plaques were visualized after 10 days. All strains exhibited similar overall fitness in vitro (representative plaques are shown in panel D). The area of 30 plaques from each parasite line was measured, and no significant difference (P > 0.05) was determined by one-way ANOVA. ns, not significant (E).