Figure 3.

Effects of olorofim exposure on S. brasiliensis yeasts. Treated yeasts analyzed by fluorimetry exhibited a dose−dependent DNA accumulation (A), increased chitin (B), decreased β−glucan (C), increased mannan (D), and reduced electronegativity (E) and conductance (F). Cell wall measurements revealed that olorofim treatment increased the thickness of this structure (G). Scanning electron microscopy images showed untreated yeasts with an elongated shape (Hi), and treated cells with alterations in cell wall integrity, exhibiting a granular appearance (arrow in Hii). Transmission electron microscopy images illustrated untreated cells with homogeneous cytoplasm with nucleus (n) and mitochondria (m), surrounded by a plasma membrane and cell wall (cw) (Hiii), while treatment with olorofim induced an amorphous shape and modifications in the cell wall structure (arrow in Hiv). Graphs showed the min and maximum with the line in the median, with the X−axis representing olorofim concentrations in µM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns: not significant by one-way ANOVA. Scale bars: 2.5 µm (Hi,Hii) and 0.5 µm (Hiii,Hiv).