Clotrimazole

Deletion and GFP-fusion strains were obtained by homologous recombination of the PCR product with heterologous selection markers, as listed in Table 1. All strains in this study are derivatives of the W303 strain. As templates for PCR, the DNA of yeast strains from Euroscarf collection, pUG27 (Gueldener et al., 2002 (link)) and pFA6a-His3MX6 (Longtine et al., 1998 (link)) plasmids were used. All newly generated strains were verified by PCR with independently designed primers (Table 2) and RT-qPCR (Supplementary Text S1, Table S1, Figure S1). We used standard yeast-rich and synthetic mediums described by Sherman (2002) (link). We obtained yeast extract from BD and D-glucose from Helicon. Clotrimazole, miconazole, nigericin, amphotericin B, propanol, butanol, and FM4–64 were obtained from Thermo Fisher Scientific. Bacto Agar, peptone, NaCl, KCl, and NaN₃ were obtained from Amresco, and 2-Deoxy-D-glucose was obtained from Chem-Impex Int’l Inc.

1,1,1-Tris(hydroxymethyl)propane
(Sigma-Aldrich) was dissolved in acetone and precipitated by addition
of diethyl ether. Then, 60 wt % NaH in mineral oil (Merck) was washed
with dry tetrahydrofuran (THF) to remove oil and dried. The oil was
removed by washing with dry tetrahydrofuran, and then drying under
reduced pressure. Benzyl glycidyl ether (BGE, Sigma-Aldrich) was dried
over CaH₂ and then distilled under reduced pressure before
use.
d,l-1,2-Isopropylidene glyceryl glycidyl
ether (IGG) was synthesized according to the procedure described by
Wurm et al.^{33 (link)} THF (Sigma-Aldrich) was dried
over a Na/K alloy. Dimethyl sulfoxide (DMSO, Sigma-Aldrich) was purified
and dried according to the procedure described in ref (34 ) and stored over 4 Å
molecular sieves.
Nifuratel (TCI) and clotrimazole (Thermoscientific)
were used as
received without any further purification. A Strat-M membrane was
used as a nonanimal model for transdermal diffusion testing to predict
the diffusion of drugs through human skin.
The simulated vaginal
fluid (SVF) of pH = 4.2 was prepared according
to the composition described in ref (35 (link)).