FIG 5.

Ergosterol quantification for UPC2A disruptants in an SDD clinical isolate, SM1 (A, C, and E), and matched resistant isolate SM3 (B, D, and F). Genotypes studied were as follows: wild-type UPC2A allele (solid circles), Δupc2a (open squares) (A and B), Δupc2b (open triangles) (C and D), and Δupc2a Δupc2b (open diamonds) (E and F). The results for the UPC2A disruptants shown in panels A, B, E, and F showed statistically significantly lower ergosterol content than was found in the wild type (P < 0.05). Knockout strains with the wild-type allele complemented back in showed no difference in ergosterol content compared to those of the wild-type isolates (data not shown). Strains were grown in RPMI medium for 16 h, followed by a heptane extraction and spectrophotometric scan between 240 and 300 nm (41). Each isolate was extracted and analyzed in three independent experiments.