Increasing the Fungicidal Action of Amphotericin B by Inhibiting the Nitric Oxide-Dependent Tolerance Pathway
Amphotericin B induced cell cycle arrest in the G2/M phase in yeast. Exponential yeast cultures were treated with either control (1% DMSO; 10% mQ), 200 mM L-NAME (dissolved in mQ), 10 μM AmB (dissolved in DMSO), or a combination of the above for 7.5 min (a) and 15 min (b). After treatment, cells were washed with PBS, fixed in 70% EtOH, stained with PI, and subjected to flow cytometry for cell cycle analysis. White bars represent cells in the G0/G1 phase, black bars represent cells in the S phase, and pixelated bars represent cells in the G2/M phase. Means and standard error of the means (SEMs) of 3 independent biological experiments () are presented. Two-way ANOVA followed by Tukey multiple comparison test was performed to analyse differences between the cell cycle distributions of control treatment and AmB, L-NAME, or AmB + L-NAME treatment and between cell cycle distributions of AmB treatment and AmB + L-NAME treatment. ,,, and represent ,,, and , respectively. Multiplicity-adjusted values are presented in the main text.