Fig. 1: Functional genomic screening in C. albicans reveals genes differentially required for normoxic and extreme hypoxic fitness.

a Schematic of the screening and analysis pipelines of GRACE libraries^18,19. DS^AC or DS^EHC cut-offs for each step are provided in the “Methods”. Genes specifically required for EHC (extreme hypoxic condition) or AC (aerobic condition) fitness are highlighted in green or purple, respectively. b Growth of homozygous deletion strains were compared with wild-type (WT) parent by a spot dilution assay on YPD and SD agar. Homozygous gene deletion is denoted by italicized gene name in lower-case through this study. c Schematic of fermentation pathway and spot dilution assay showing adh1 and pdc11 mutants are blocked in EHC growth. d WT and adh1 mutant strains grown in liquid SD medium under EHC were sampled at indicated time points. Whole cell ATP levels and medium ethanol concentration were normalized to biomass determined by optical density at 600 nm (OD). LCD Limit of confident detection. e Spot dilution assay of hit strains. An overnight culture of each strain grown in SD ± 0.05 μg mL⁻¹ DOX were serially diluted and spotted on SD agar ± 20 μg mL⁻¹ DOX, respectively. Media were supplemented with 20 mg L⁻¹ histidine. f AC and EHC growth of mutant strains was compared as described in (b). g, h, DOX treatment depletes tetO-driven HSF1 expression under EHC. Overnight cultures were grown in YPD ± 0.1 μg mL⁻¹ DOX were respectively plated on YPD ± 20 μg mL⁻¹ DOX agar and grown for 24 h under EHC before analyzed by (g) RT-qPCR or (h) anti-TAP immunoblotting. In (h), loading of the non-DOX-treated sample was titrated. The blot was probed for Hsp90 to show that Hsp90 was not depleted with Hsf1. CBS Coomassie Blue Staining for loading control. d, g show mean ± standard deviation (SD) of biological triplicates. Unpaired t-tests were performed and two-tailed p-values reported.