FIGURE 3.

Lack of NcGrx1 adversely affects the growth of parasites. (A) Strategy for construction of ΔNcGrx1 and ΔNcGrx3 strains. (B) PCR identification of ΔNcGrx1 and ΔNcGrx3 parasites using the primers indicated above. (C) Plaque assays comparing the growth of wild-type, knockout, and overexpression parasites. Each well was infected with 300 parasites, and plaques were stained for 9 days. The plaque areas were counted by randomly selecting at least 20 plaques and measured using the pixel point in Photoshop C6S software (Adobe, United States). The data were compiled from three independent experiments. (D) Quantification of plaque area sizes. Analysis of the plaque area was performed using t-test. (E) Mouse survival after infection with different strains. BALB/c mouse (n = 5) were injected intraperitoneally with 8 × 10⁶ doses of parasites. Statistical analysis was performed using survival curve of Graph Pad Prism (SAS Institute Inc., United States). ***p < 0.001.