Fig. 4

Adjuvant drugs significantly reduce FLC tolerance but not resistance and render FLC cidal. a Disk diffusion assays performed with 25 µg FLC on casitone plates supplemented with adjuvant drugs 20 µg/ml fluoxetine, 5 ng/ml aureobasidin A, 0.5 ng/ml rapamycin, 10 µg/ml fluphenazine, 12.5 ng/ml staurosporine, 0.25 µg/ml tunicamycin, Hsp90 inhibitors (0.5 µg/ml geldanamycin and 0.5 µg/ml radicicol), and calcineurin inhibitors (0.5 µg/ml FK506 and 0.4 µg/ml cyclosporine A) shown for strain SC5314. b RAD and FoG levels performed on disk diffusion assays with FLC and adjuvants. c Effect of drug adjuvants and pathways inhibitors on the viability of cells growing inside the zone of inhibition. FLC disk diffusion assays of SC5314 without or with adjuvant were replica plated (after removal of the drug disk) onto casitone plates (without FLC or adjuvants) and incubated at 30 °C for 48 h. d, e FLC disk diffusion assays performed using a series of mutants carrying deletions in genes encoding the calcineurin subunit Cnb1, the calcineurin-responsive transcription factor Crz1, calcineurin regulators Rcn1 and Rcn2, MAP kinase Mkc1, vacuolar trafficking protein Vps21 (d), ergosterol biosynthesis regulator Upc2, and efflux pump regulators Tac1 and Mrr1 (e). These mutants as well as the rcn1 crz1 double mutant were analyzed by diskImageR, and RAD and FoG levels are shown relative to the isogenic parental strains (WT). All pictures in this figure are representative of two biological replicates, asterisks denote significant differences relative to corresponding parental strains. For all panels, n ≥ 2; ***P < 0.001; **P < 0.01; *P < 0.05, error bars denote standard deviations