Parameter | Cell line | Mean | SD | P value^{d} |
---|---|---|---|---|

% infected cells^{b} | ||||

24 h | WT | 84.60 | 9.80 | *** |

Cas9/T7 | 80.39 | 8.43 | ** | |

4E-3(+/−) | 49.73 | 7.75 | ||

Addback | 84.41 | 9.89 | ** | |

4 h | WT | 95.79 | 5.43 | ** |

Cas9/T7 | 87.50 | 6.66 | ** | |

4E-3(+/−) | 39.32 | 10.15 | ||

Addback | 96.30 | 4.79 | ** | |

No. of parasites per infected cell^{c} | ||||

4 h | WT | 4.68 | 1.50 | ** |

Cas9/T7 | 3.95 | 1.06 | ** | |

4E-3(+/−) | 1.58 | 0.40 | ||

Addback | 4.53 | 1.16 | ** | |

24 h | WT | 12.60 | 2.82 | *** |

Cas9/T7 | 11.90 | 1.07 | *** | |

4E-3(+/−) | 2.14 | 0.28 | ||

Addback | 13.96 | 4.39 | *** |

↵a The ability of the LeishIF4E-3(+/−) mutant to infect the macrophage RAW 264.7 cells was quantified by counting the number of infected macrophages. Control infections included wild-type parasites, the Cas9/T7-expressing cells, and the LeishIF4E-3 addback cells, in which expression of LeishIF4E-3 was recovered.

↵b Infectivity of each cell line was monitored by confocal microscopy as described in the legend to Fig. 6, at 24 and 4 h postinfection.

↵c Parasite loads in infected macrophages were also determined, by counting the number of parasites per infected macrophage. The cells were counted using a cell counting plugin of Image J. A total of 100 macrophages were counted three times for each parasite cell line.

↵d The statistical analysis for percentage of infected cells and average parasite per cell, along with standard deviation values (SD), was performed by Kruskal-Wallis test using GraphPad Prism. The number of infected macrophages and average number of parasite counts per infected macrophage were the two parameters that were compared between LeishIF4E-3(+/−) deletion mutants and each of the control lines: wild type, transgenic cells expressing Cas9/T7, and LeishIF4E-3 addback cells. ***,

*P*< 0.001; **,*P*< 0.01. The statistical difference between the control lines was nonsignificant.