Identification via microarray analyses of selective mRNAs that accumulate in the nucleus as a result of fragile X premutation rCGG repeats.
A. The three sets of microarray experiments were carried out in triplicate (three biological replicates) with equal amounts of total RNA obtained from the Drosophila heads of control total, nuclear, and cytoplasmic fractions and the corresponding experimental premutation rCGG repeat samples of total, nuclear, and cytoplasmic fractions. Microarray analyses were carried out by identifying significantly changed genes at the 0.001 level of the univariate test. B. Scatter plot of mean log intensities of each sample demonstrating differentially expressed significant genes with fold-change of 2 or more between the classes within a particular set. C. The numbers of unique differentially expressed genes generated by the different cellular compartments of rCGG sample (comparison of total, cytoplasmic, and nuclear fraction) are displayed in Venn diagrams. D. Fold enrichment depicted by ratios between the intensities of normalized log-transformed gene expressions for 45 genes unique to CGG nuclear fractions in various classes is displayed using the Cluster and TreeView programs for the wild-type and premutation rCGG datasets. The fold of the change is indicated on both sides of the scale bar E. Validation of nuclear enrichment by real-time PCR analysis of the selective genes. Real time against Fmr1 serves as a control. The data represent mean ± SEM, n = 3.