Fold changes were calculated

for day 2 spherules vs mycelia and day 8 spherules vs mycelia. For each gene, the absolute peak log 2 fold change (FC) was identified across the three conditions and the raw expression values for the top 100 were log transformed and median-centered and included in the heatmap. Hierarchical clustering of genes and array samples based on their expression profiles is reflected in the dendrograms to the left and the top of the heatmap respectively and was performed selleck products by calculating distances using the Pearson correlation metric and then clustering distances using the average linkage method. The expression of genes marked with an asterisk (*) was confirmed by RT-qPCR. The scale is shown: red shading indicates greater expression blue shading represents lesser expression. Figure 3 Venn diagrams showing the number of genes that are differentially expressed in day 2 spherules and day 8 spherules compared to mycelia. The

number of up- or downregulated genes in shown. The procedures for determining up- or downregulation are in the methods section. There were a total of 2208 genes (22% of the genome) that were differentially expressed between spherules at one or both the time points we studied and mycelia. Figure  4 shows Venn diagrams depicting up- and downregulated genes in day 2 and day 8 spherules compared to mycelia. About a third of the differentially expressed

genes were up- or downregulated in both day 2 and day 8 spherules compared to mycelia. Florfenicol However, similar numbers of genes were exclusively upregulated in either selleck kinase inhibitor day 2 (N = 443) or day 8 (N = 319) spherules, or exclusively downregulated at either day 2 (N = 565) or day 8 (N = 233) spherules. The difference in gene expression between day 2 and day 8 spherules was apparent when we compared day 2 and 8 spherules directly to each other; 1,197 differentially expressed genes (12% of the total genome) were identified (Additional file 4: Table S2). Therefore, although gene expression by environmental form of the fungus and the parasitic form were quite distinct as might be expected, gene expression by young and mature spherules was also quite different from each other. Not only were there differences in which genes were expressed at each stage, but also the degree of modulation was large. For example, the maximum difference in expression of a gene (CIMG_10264) between day 2 spherules and mycelia was 48.6 fold and the median modulation between mycelia and day 2 spherules was 3.26. Figure 4 Confirmation of gene expression differences by RT-qPCR between day 2 spherules vs mycelia, day 8 spherules vs mycelia and day 8 vs day 2 spherules. The figure shows a comparison between the fold change for each gene for RT-qPCR data (grey bars) and microarray data (black bars) between the different conditions.