Journal: G3: Genes|Genomes|Genetics
Article Title: Php4 Is a Key Player for Iron Economy in Meiotic and Sporulating Cells
Figure Lengend Snippet: Transcriptomic response of S. pombe as a function of changes in iron levels and effect of php4 ∆ /php4 ∆ disruption relative to wild-type cells during meiotic differentiation. (A) Cultures of pat1-114/pat1-114 php4 + /php4 + cells were precultivated in the presence of Fe (0.74 µM) and Dip (50 µM) for 16 hr. Pretreated cells were further exposed to Fe (100 µM) and Dip (75 µM), respectively, and then immediately induced to undergo synchronous meiosis. After 7 hr of meiotic induction, total RNA was extracted and used for microarray experiments. The graph represents a genome-wide picture of differentially expressed genes (X axis) in iron-replete vs. iron-limited cells. For simplicity, only a few differentially expressed transcripts are labeled on the graph. (B) Genome-wide picture of differentially expressed genes in pat1-114/pat1-114 php4 Δ /php4 Δ vs. pat1-114/pat1-114 php4 + /php4 + cells that had been precultured (50 µM) and cultured (75 µM) in the presence of Dip. Genome-wide transcripts (X axis) were analyzed by DNA microarrays. All differentially expressed genes are depicted, although only a few of them are labeled on the graph for the sake of clarity. (C) Venn diagram representing the number of genes that were induced in iron-treated php4 + /php4 + cells compared to php4 + /php4 + cells grown under iron-limiting conditions and the number of genes that exhibited higher expression levels in iron-starved php4 ∆/ php4 ∆ compared to php4 + /php4 + cells grown under the same conditions. Dip, 2,2’-dipyridyl; Fe, iron(III) chloride.
Article Snippet: First, the microarray gasket slide from Agilent Technologies was improved in that it contained a larger number of probes (15,000) which allowed a ∼2–3 × increased coverage for each S. pombe ORF.
Techniques: Microarray, Genome Wide, Labeling, Cell Culture, Expressing