IL-36R stimulation of colonic mouse fibroblasts. IL-36R stimulation of colonic mouse fibroblasts

Intestinal fibroblasts from the colons of C57BL/6 wild-type mice were grown in the presence or absence of IL-36ÉŁ (30 ng/ml) for 4 hours. The experimental setup included 6 samples from 3 colons in a pairwise design (3 stimulated vs 3 unstimulated). Profiling of whole-genome transcriptomic patterns was performed with 1.2 Âľg high quality total RNA per sample by RNA-seq at the Next Generation Sequencing core unit of the university hospital Erlangen. The sequencing platform was Illumina HiSeq-2500 with high output mode and single-end 100bp fragments. The sequencing library was NuGEN Ovation RNA-Seq Library Prep-Kit. Demultiplexed reads were corrected regarding rRNAs, tRNAs, mt-rRNAs and mt-tRNAs. Alignment against the mus musculus reference genome (Ensemble version 75 for GRCm38) was performed with RNAseq aligner STAR (version 2.4.0.i) and quantification (HTseq count) was done of unique mappings.