Suloctidil

Apogossypol and Leflunomide from ApexBio (Boston, MA, USA), A-1331852, A-1210477, ABT-199, Z-VAD.FMK and CCCP from Selleck (Houston, TX, USA), Teriflunomide, norhydroguaiaretic acid (NDGA), Ivermectin, Terfenadine, Suloctidil, orotate and uridine from Sigma Aldrich (St Louis, MO, USA), MitoTracker Deep Red FM from Thermo Fisher (Loughborough, UK) were used. Antibodies against BAP31, RTN4, BiP, PDI, CHOP, DHODH and tubulin from Abcam (Cambridge, UK), CLIMP-63 and BAX (6A7) from Enzo Life Sciences (Exeter, UK), TIM22 and KNT-1 from Sigma, HSP60, Cytochrome c, BAX, OPA1 and DRP-1 from BD Biosciences (San Jose, CA, USA), phospho-DRP-1 (S616), phospho-DRP-1 (S637), MFN1 and MFN2 from Cell Signaling Technologies (Danvers, MA, USA), BAK (AB-1) from Calbiochem (Watford, UK), MFF, MID49 and MID51 from ProteinTech (Manchester, UK) and GAPDH from Santa Cruz Biotechnologies (Santa Cruz, CA, USA) were used. For RNA interference, cells were transfected with 10 nM of siRNAs against DHODH (SI00363384 and SI00363391) purchased from Qiagen Ltd. (Manchester, UK), using Interferin (Polyplus Transfection Inc, NY), according to the manufacturer’s protocol and processed 72 h after transfection.

This study utilized wild type strains of C. albicans, YEM30 and LC3, isolated from a patient with vaginal candidiasis from QiLu hospital. All strains were stored in 20% glycerol at -80°C prior to the experiments. The frozen glycerol stock of the strain was regularly revived on YPD agar medium (1% yeast extract, 2% tryptone, 2% glucose, and 2% agar). For the broth culture, strains were grown in YPD medium at 30°C with agitation (100 rpm). RPMI-1640 medium was buffered with morpholinepropanesulfonic acid buffer (Sigma-Aldrich, St. Louis, US) to pH 7. Stock solutions of suloctidil (Sigma-Aldrich, St. Louis, US) were prepared in dimethyl sulfoxide (DMSO, Sigma-Aldrich, St. Louis, US) and stored at -20°C until further use.