RNA-Seq analysis of the pancreatic islets from 6-week-old Paupar WT and KO mice

Transcriptional analysis of the pancreatic islets of adult Paupar KO mice versus Paupar WT mice. Total RNA from Paupar WT and Paupar KO mice was converted into cDNA libraries (TruSeq RNA sample preparation kit, Illumina) according to manufacturer's instructions. Sequencing was performed to a depth of 30 million, single-end reads in three biological replicates per genotype. Reads were aligned to the mouse genome (mm9) using HISAT2 v2.1.0 (Kim et al., 2013). Aligned reads were assigned to genes using annotations from Ensembl (Mus_musculus.GRCm38.73.gtf) and HTseq-count v0.10.0 (Anders et al., 2015). Differential expression across cohorts was assessed using DESeq2 v1.18 (Love, Huber, and Anders, 2014). Among the downregulated genes with a p-value < 0.05 genes important for alpha cell identity and function. Overall design: mRNA profiles of the pancreatic islets of 6 week old Paupar WT and KO mice were generated by deep sequencing, in triplicate, using Illumina HiSeq2000.