FIG 2

SNP+SHAM treatment leads to cell wall alterations and altered lipid metabolism. (A) Wild-type cells were serially diluted (1:10 dilutions), and equal volumes were spotted onto YPD plates (at 30 or 37°C) containing 1 mM SNP–0.5 mM SHAM and 25 µg/ml calcofluor white (CFW) or 40 µg/ml Congo red (CR). (B) Cells were treated with 1 mM SNP–0.5 mM SHAM for 18 h and processed for transmission electron microscopy (TEM). Representative examples of untreated and SNP+SHAM-treated cells are shown. Bar = 100 nm. (C) Quantification of outer cell wall (o) and inner cell wall (i) thicknesses was carried out from TEM images. Measurements were taken at 5 points along the cell wall for 25 cells per group. Bar = 100 nm. (D) Cell wall material was extracted and acid hydrolyzed from cells treated with 1 mM SNP–0.5 mM SHAM for 18 h, followed by HPLC analysis (n = 3). Graphs show means ± standard deviations. (E) TEM images from cells treated with SNP+SHAM. Arrows indicate lipid droplets. Bar = 500 nm. (F) Control and SNP-, SHAM-, or SNP+SHAM-treated cells were stained using LD540 neutral lipid stain and viewed by fluorescence microscopy. Bar = 10 µm. Student's t test was used to compare groups. *, P < 0.01.