RNAseq gene expression profile of pancreatic islets from mBD14-treated NOD mice (Miani et al., CELL-METABOLISM-D-18-00033)
RNA-seq analysis of pancreatic islets from 10-wk-old female NOD mice treated with mBD14 (10 µg/mouse) or vehicle (d‒5). Methods: Total RNA were isolated from pancreatic islets (>150) using the RNeasy Kit (QIAGEN) including a DNAse treatment step. RNA quality was assessed using RNA Screen Tape 6000 Pico LabChips with the Tape Station (Agilent Technologies) and RNA concentration was measured by spectrophometry using the Xpose (Trinean). RNAseq libraries were prepared starting from 1 µg of total RNA using the TruSeq Stranded mRNA LT Sample Prep Kit (Illumina) as recommended by the manufacturer. Half of the oriented cDNA produced from the poly-A+ fraction was PCR amplified (11 cycles). The RNAseq libraries were sequenced on an Illumina HiSeq2500 (Paired-End sequencing 130x130 bases, High Throughput Mode). A mean of 23 million of paired-end reads was produced per library sample (between 21 to 25 million of passing filter reads). The generated data were analyzed using the Ingenuity Pathway Analysis software (Qiagen).