FIG 3.

sre1Δ and rim101Δ mutant strains have varied changes in cell wall chitin exposure and interactions with host immune cells. (A) Staining of rim101Δ, sre1Δ, and wild-type cells with calcofluor white (CFW) and wheat germ agglutinin (WGA). Cells were incubated in CO₂-independent medium for 18 h at 37°C. Cells were stained with FITC-conjugated WGA and CFW and incubated in the dark for 35 min and 10 min, respectively. Mean fluorescence intensity was quantified for each strain and each condition. Two-way ANOVA and Tukey’s multiple-comparison test were run to determine statistical significance. White scale bars indicate 5 μm. ****, P value < 0.0001. DIC, differential inference contrast. (B) When grown in the presence of J774A.1 macrophages, the rim101Δ mutant strain can survive significantly better than both the wild-type and the sre1Δ mutant strain. Indicated strains were coincubated with macrophages for 24 h, and survival was determined by quantitative cultures. One-way ANOVA and Tukey’s multiple-comparison tests were run to assess statistical significance between fungal cell survival percentages. Six biological replicates of each strain were analyzed. **, P value < 0.003; ***, P value < 0.0002.