Figure 7
Hypoxia affects transcript levels of enzymes involved in glycolysis consistent with fermentation and cell wall components. A. KEGG pathway heat map representation of genes involved in glycolysis. Microarray datasets include three biological and two technical replicates. Microarrays compare wild type Aspergillus fumigatus strain CBS144.89 at the indicated times after exposure to hypoxic conditions to the time point immediately prior to hypoxia exposure (0 hours) using the median value from all replicates. Yellow indicates transcript level is higher in hypoxia. Scale indicates degree of change. B. RT-PCR of Glyceraldehyde-3-phosphate-dehydrogenase supported the observation that this was one of the most abundant hypoxia responsive transcripts in the microarray, as well as in the proteomics analysis. C. Six genes in the glycolysis pathway in Aspergillus fumigatus. Transcript levels of 6-phosophofructokinase, hexokinase A and pyruvate kinase were not dramatically altered. Transcript levels of phosphoglycerate kinase and glucokinase A are 2 to 4-fold increased in response to hypoxia. Aldehyde dehydrogense A transcript level was significantly decreased at 6, 12 and 24 hours. All RT-PCRs were performed on BioRad MyIQ real-time PCR detection system with IQ SYBR green supermix. The ΔΔC t method was used to combine all biological and technical replicates for each transcript, using β-tubulin as the housekeeping gene for comparison. D. Heat map representation of cell wall component transcripts that were compiled from literature [36]. Both microarray datasets include three biological and two technical replicates, and compare wild type Aspergillus fumigatus strain CBS144.89 at the indicated times after exposure to hypoxic conditions to the time point immediately prior to hypoxia exposure (0 hours) using the median value from all replicates. Yellow indicates transcript level is higher in hypoxia.