Journal: PLoS Genetics
Article Title: Interferon Regulatory Factor 8 Regulates Pathways for Antigen Presentation in Myeloid Cells and during Tuberculosis
doi: 10.1371/journal.pgen.1002097
Figure Lengend Snippet: BMDMs RNA was obtained from individual wt and IRF8 mutant F2 mice either prior to (unstimulated control) or 3 hrs following stimulation with IFNγ/CpG (6 samples per experimental group; 24 samples in total), and hybridized to microarrays. (A) By using a 2×2 interaction Anova analysis, 368 genes were recognized to be significantly differentially modulated by IRF8 in response to IFNγ/CpG exposure between the wt and IRF8 mutant cells. The expression profiles are ordered by hierarchical clustering; the genes, illustrated by their specific signal intensities (Log 2 scale) are displayed as rows and individual mouse samples/conditions as columns. Red coloring signifies high level of expression; green coloring denotes low level of expression. The dendrogram illustrates the clustering of the samples according to expression pattern similarities. RNA samples (6 per experimental group) used for this transcriptional profiling analysis were pooled and used for qPCR validation. Ephx1 , Cyp27a1 , Ciita , and Il10ra were selected as genes positively affected by the presence of functional IRF8 following IFNγ/CpG exposure (B), while Ms4a7 , C1qb , Angptl4 , and Slc40a1 were selected as genes negatively affected by the presence of a functional IRF8 following IFNγ/CpG exposure (C). The ratios of expression (IFNγ/CpG-stimulated versus unstimulated control), represented by fold induction, were calculated for the wt and IRF8 mutant mice separately (white bars), and compared to the corresponding microarray results (black bars). The black dots indicate the qPCR values obtained for each replicate. The microarray results were statistically significant according to Anova analysis ( t test p value of 0.05 and a fold-change cutoff of 1.5X). Hprt was used to standardize the mRNA levels of target genes for qPCR.
Article Snippet: The GeneSifter™ microarray data analysis system (Geospiza Inc., Seattle, WA, USA) was used to examine data generated from comparisons between control (unstimulated) and IFNγ/CpG-stimulated (3 hrs) groups.
Techniques: Mutagenesis, Expressing, Functional Assay, Microarray
