Dihydroartemisinin

All C. glabrata strains used in this study are listed in Table S1. The mutant strains were constructed by standard genetic techniques under a C. glabrata ATCC 55 background, and knockout strains were constructed by homologous recombination following a previously described protocol [19 (link)]. Yeast strains were cultivated in YPD medium (1% yeast extract, 1% peptone, 2% glucose) or YPG medium (1% yeast extract, 1% peptone, 2% glycerol); solid media contained 1.5% agar. FDA-approved drugs used in screening antifungal drugs were purchased from Selleck (Houston, TX, USA), this library included artemisinin and its analogues artesunate, dihydroartemisinin, artemether.

The Pathogen Box was provided by the Medicines for Malaria Venture (MMV) and was comprised of 10 mM solutions in dimethyl sulfoxide (DMSO) of each of 400 compounds. Details of the compounds in the Pathogen Box are set out at CHEML-NTD (https://www.ebi.ac.uk/chemblntd; accessed 20^th April 2017) and the MMV Pathogen Box website (https://www.pathogenbox.org/; accessed 20^th April 2017). Each of the 10 mM solutions was diluted ten-fold in DMSO to yield 1 mM stocks which were dispensed as 10 µL aliquots into the individual wells of a 96-well plate for storage. The PfATP4-associated antimalarial KAE609 (cipargamin) and additional quantities of the Pathogen Box compounds MMV020136, MMV020710, MMV020520, MMV006239, MMV000858, MMV020391, MMV020081, MMV001059, MMV020623, MMV688980 and MMV085210 were obtained from MMV. Dihydroartemisinin was purchased from SelleckChem and chloroquine and artemisinin were purchased from Sigma. The MMV compounds, artemisinin and Dihydroartemisinin were dissolved in DMSO; chloroquine was dissolved in water. All compound solutions were stored at −20 °C and thawed immediately prior to experimentation. Stock solutions were subjected to a maximum of 5× freeze-and-thaw cycles before being discarded.