An IL-23R/IL-22 circuit regulates epithelial serum amyloid A to promote local effector Th17 responses [RIP-Seq]. Mus musculus

Purpose: The goals of this study are to compare RNAs bound by translating ribosomes in the presence or absence of Segmented Filamentous Bacteria (SFB) colonization of the small intestine Methods: EDTA fractions containing small intestine epithelial cells from SFB negative and SFB positive mice were harvested. TRAP assay enriching for ribosome bound fraction were enriched using anti-GFP antibodies against GFP-ribosomeL10 fusion protein. mRNA profiles were generated by deep sequencing, using Illumina HighSeq. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) and TopHat followed by CuffDiff. qRT–PCR validation was performed using SYBR Green assays. Results: SFB colonization upregulate a subset of mRNAs being translated by ribosomes. Conclusions: SFB changes small intestine epithelial cell translation profile. Overall design: compare RNAs bound by translating ribosomes in the presence or absence of Segmented Filamentous Bacteria (SFB) colonization of the small intestine