Increased cell wall chitin and β-glucan in <i>A. fumigatus</i> isolate Af5517.

<p>(<b>A,B</b>) Representative dot-blot images of processed <i>A. fumigatus</i> mycelial extracts and chitin (shrimp shell chitin) (<b>A</b>) or β-glucan (curdlan) (<b>B</b>) standards, probed with chitin binding probe or anti-(1,3)-β-glucan, respectively. Blots of mycelial extracts depict 75 µg (A) and 100 µg (B) of total protein of each isolates. (<b>C,D</b>) Chitin (<b>C</b>) or β-glucan (<b>D</b>) vs. total fungal protein in mycelial extracts as determined by dot blot assay. (<b>C</b>) Chitin was significantly increased compared to other isolates at 25, 50, or 75 µg of total protein, while β-glucan (<b>D</b>) was significant at 50 and 100 µg total protein. ****p<0.0001. (<b>A–D</b>) Panels are representative of two experiments. (<b>E,F</b>) Representative flow cytometric histograms of dormant (0 h) or swollen (5 h at 37°C) conidia of each isolate stained with the chitin binding wheat germ agglutinin (WGA, panel E) or anti-(1,3)-β-glucan (<b>F</b>). Negative controls are unstained conidia (<b>E</b>) or goat anti-mouse alexa-fluor 488 only (<b>F</b>) and depicted as dotted histograms. (<b>G,H</b>) Median fluorescence intensities of WGA (<b>G</b>) or anti-β-glucan (<b>H</b>) stained conidia after 0 or 5 hours incubation at 37°C. (<b>G</b>) Chitin exposure in Af5517 conidia was significantly increased in comparison will all other isolates after 0 and 5 hours incubation. *p<0.05. **p<0.01. Panels are a summary of three experiments.