Figure 2: Haploinsufficiency (HIP) profiling identifies potential targets of GW461484A (GW).

A, A pooled library of double bar-coded heterozygous deletion mutants covering ~90% of the C. albicans genome was grown in the presence or absence of GW (3 μM). Genomic DNA was isolated, and the up and down strain-specific barcodes (UP-TAG and DOWN-TAG) were PCR-amplified, pooled, and high-throughput sequencing was performed to assess the relative abundance of each tagged strain. Strains were considered significantly reduced in abundance if the solvent/drug log₂ ratio was greater than four median absolute deviations (MADs) above the median for both the UP-TAG and DOWN-TAG, or if either the UP-TAG or DOWN-TAG was >4MAD and the opposing TAG was omitted due to low total reads. Significantly enriched UP-TAGs and DOWN-TAGs are shown in blue and orange, respectively. Grey dots represent UP-TAGs and DOWN-TAGs of strains for which only the UP-TAG or DOWN-TAG was significant, and thus the strain was not classified as a hit, as well as strains where neither tag was significant. B, Functional annotation of genes for which heterozygous deletion causes significant hypersensitivity to GW. Gene ontology (GO) terms were identified using the GO component finder available on the Candida Genome Database. Significantly enriched GO terms are indicated above each cluster of colored boxes highlighting individual genes (*** p<0.000005, **p<0.005). Some genes were shared across GO terms but are displayed only in the cluster with the greatest statistical significance. Red outlines indicate genes that encode kinases. (See also Figure S2).