Figure 8.

Yeasts present in the granuloma produces low levels of intracellular siderophore ferricrocin and has low lipid availability. For the ferricrocin levels analysis (A) a quadruplicate of lung granuloma, control lung, control yeasts and recovered yeasts were frozen, and 100 mg of frozen material was macerated with the help of a pestle and pot. The secondary metabolites extraction was performed with 1 mL of methanol (100%; HPLC grade), followed by sonication bath for 40 minutes. For sample preparation, 500 μL of each sample extract were filtered in a 0.22 μm PTFE and diluted to 1 mL volume with HPLC-grade methanol. Then LC-HRMS/MS analysis were performed and data were processed with Xcalibur software (version 3.0.63). Statistical analysis was performed using one-way ANOVA. Bars represent means ± standard error of the feature area of groups of 4 to 5 samples (*p<0.05; **p < 0.01). For lipid availability analysis (B) control yeasts, granuloma yeasts and recovered yeasts were stained with filtered Oil Red O for 2 hours at room temperature. Subsequently, the cells were washed twice with ultrapure water and then resuspended in PBS for analysis on a microscope slide. The photos were taken using the Leica DM750 microscope and the Leica Application Suite V4.8 software, thus allowing the observation of stained neutral lipids inside the cells.