, two and four hours and total RNA was isolated followed by qRTPCR (See
, 2 and four hours and total RNA was isolated followed by qRTPCR (See Supplies and Approaches for details). As anticipated, expression of SFLTAP steadily improved from time points 0 h to 4 h (Figure 5A, left panel). This improved SFL expression correlated with decreased SFL2 and BRG expression (Figure 5A, middle and suitable panels), constant having a unfavorable regulation of SFL2 andPLOS Pathogens plospathogens.orgBRG expression. However, PPCKinduced SFL2TAP expression (Figure 5B, left panel) correlated with decreased expression of SFL (Figure 5B, SFL panel) and elevated expression of UME6 and ALS3 (Figure 5B, UME6 and ALS3 panels), consistent with our microarray information (Figure four). Taken together, our transcriptomics data reflect the antagonistic functions of Sflp and Sfl2p in regulating C. albicans morphogenesis, with SFL advertising the yeastform growth which correlates with upregulation of yeast formspecific genes and downregulation of HSGs, and SFL2 promoting hyphal growth which correlates with upregulation of HSGs and downregulation of yeast formspecific genes.The Sflp and Sfl2p NK-252 custom synthesis regulatory networkWe combined the transcriptomics as well as the ChIPSeq data to be able to get a genomewide view with the transcriptional modules associated with Sflp and Sfl2p regulatory functions (Figure six). We had been expecting to PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/24121451 discover a substantial quantity of genes which might be bound by Sflp and downregulated in pCaEXPSFLHA3 vs. pCaEXP microarray data, as Sflp is thought to act as a repressor. In line with all the function of Sfl2p as an activator of hyphal development, we were also hypothesizing that binding of Sfl2p to its targets would correlate with elevated expression of those target genes.C. albicans Sflp and Sfl2p Regulatory NetworksFigure six. Sflp and Sfl2p transcriptional modules. Venn diagrams on the overlap amongst the genes that happen to be modulated in (A) SFL or SFL2 transcriptomics (light red circles, upregulated; light green circles, downregulated; gene expression foldchange cutoff .5; Pvalue cutoff 0.05) and normally bound by Sflp and Sfl2p (light blue circle) or (B) SFL2 transcriptomics (light red circle, upregulated; light green circle, downregulated;PLOS Pathogens plospathogens.orgC. albicans Sflp and Sfl2p Regulatory Networksgene expression foldchange cutoff .5; Pvalue cutoff 0.05) and especially bound by Sfl2p (light grey circle). Numbers in the Venn diagrams indicate the amount of genes. Circled numbers indicate the number of genes which can be (A) both modulated in SFL or SFL2 transcriptomics information and typically bound by Sflp and Sfl2p or (B) each modulated in SFL2 transcriptomics data and especially bound by Sfl2p. The name of those genes (or their orf9 nomenclature) along with the functional categories to which they belong are shown inside the linked boxes. , DCK is needed for hyphal formation; orf9.3475 can be a hyphal induced gene. doi:0.37journal.ppat.00359.gSurprisingly, amongst the three targets usually bound by Sflp and Sfl2p, 40 genes had 
been upregulated and only 22 genes were downregulated in pCaEXPSFLHA3 vs. pCaEXP information (Figure 6A). Conversely, 39 genes have been downregulated in pCaEXPSFL2HA3 vs. pCaEXP information and only five genes were upregulated (Figure 6A), indicating that Sflp and Sfl2p have dual transcriptional regulatory functions; acting as both transcriptional activators and transcriptional repressors. As Sflp and Sfl2p respectively act as a repressor and an activator of hyphal development, we examined the set of genes that had been typically bound by these two regulators and whose exp.
