Fig. 3. CBS mediates the cell wall remodeling of C. albicans.

(A) Transcriptional variations of several cell wall– and ER-related genes in C. albicans strains cultured in SD medium for 5 hours at 30°C. (B) Phosphorylated Mkc1 and Cek1 levels in the indicated C. albicans strains within 24 hours of growth in SD medium at 30°C. (C to E) The mannan content of the indicated strains was assessed using the concanavalin A–fluorescein isothiocyanate (ConA-FITC) staining method. Representative micrographs of ConA-FITC–stained samples are shown (C). Scale bar, 5 μm. Cell wall structures were also examined using transmission electron microscopy (TEM) (C). Scale bar, 100 nm (TEM). The fluorescence intensity of ConA-FITC was quantified by flow cytometry to determine the mannan content (D), and the GMean values of fluorescence intensity were calculated (E). (F) The mannan content in C. albicans was also measured using a chemical extraction method. (G to I) Surface β-glucan exposure in C. albicans strains. Cells of the indicated C. albicans strains were stained with an anti–β-1,3-glucan monoclonal antibody, followed by staining with a tetramethylrhodamine-5-(and-6)-isothiocyanate (TRITC)–labeled secondary antibody to visualize β-1,3-glucan by confocal microscopy. Representative micrographs of β-glucan exposure are shown (G). Scale bar, 5 μm. The fluorescence intensity was quantified by flow cytometry to indicate β-glucan exposure (H), and the GMean values were calculated (I). Student’s t test (A) and one-way ANOVA followed by Tukey's test (E, F, and I) were used to determine statistical significance. *P < 0.05, **P < 0.01, and ***P < 0.001.