FIG 5.

Virulence contributions of the C. neoformans arrestin family. (A) The WT, ali1Δ mutant, and ali1Δ + ALI1 strains were coincubated with J774A.1 murine macrophages at a multiplicity of infection (MOI) of 1 for 24 h. Survival of the strains was assessed by quantitative culture, and the percentage of recovered CFU was normalized to the WT strain. This experiment was performed with six biological replicates (n = 6). Error bars represent the SEM. Log transformation was used to normally distribute the data for statistical analysis (one-way ANOVA; ns, not significant). (B) Female C57BL/6 mice (n = 10) were intranasally inoculated with 10⁴ CFU of the WT, ali1Δ mutant, or ali1Δ + ALI1 strain. Mouse survival was tracked for 50 days postinfection (log rank test). (C) The WT strain, the MATa KN99 strain, and three isogenic but independent arrestin null mutants (all also MATa) were coincubated with J774A.1 murine macrophages at an MOI of 1 for 24 h. Survival of the strains was assessed by quantitative culture, and the percentage of recovered CFU was normalized to the WT strain. This experiment was performed with four biological replicates (n = 4). Error bars represent the SEM. Log transformation was used to normally distribute the data for statistical analysis (one-way ANOVA; *, P < 0.05; **, P < 0.01). (D) Female C57BL/6 mice (n = 10) were intranasally inoculated with 10⁴ CFU of the WT strain and a representative arrestin null mutant, ali1Δ ali2Δ ali3Δ ali4Δ-#2 (CLT57). Mouse survival was tracked for 50 days postinfection (log rank test; ****, P < 0.0001).