Defects in intracellular trafficking of fungal cell wall synthases lead to aberrant host immune recognition
(A) The mar1Δ cell wall changes are not dependent on temperature. The mar1Δ strain was incubated for 16–18 hours with shaking at the indicated temperature in rich medium (YPD) or tissue culture medium (TC). Exposed chitooligomers in the cell wall were stained with FITC-conjugated WGA and imaged by fluorescent microscopy using the GFP filter. (B) Average fluorescence of at least 100 individual cells was measured using ImageJ/Fiji software. Ns, not significant as determined by one-way ANOVA with Tukey’s multiple comparisons test; all other comparisons, p < 0.0001. (C) The mar1Δ cell wall changes occur with increased pH and can be partially suppressed by glucose supplementation. mar1Δ cells were incubated for 16–18 hours with shaking at 30°C in YPD, YPD buffered to pH 7.4, TC, or TC supplemented with 2% glucose. Cells were stained with FITC-conjugated WGA and imaged as above. Bar, 10 μM. (D) Average fluorescence was measured as above. ****, p < 0.0001; ns, not significant, as determined by one-way ANOVA with Tukey’s multiple comparisons test.