Hiseq1500 equipment

RNA was prepared with the RNeasy Plus Micro Kit (Qiagen). RNA-seq library samples were prepared using an NEBNext Ultra RNA Library Prep Kit for Illumina (New England BioLabs). The sequencing was carried out using a HiSeq1500 equipment (Illumina). The sequence reads were mapped to the mouse genome (NCBI version 37) using TopHat2 (version 2.0.8) and botwie2 (version 2.1.0) with default parameters, and gene annotation was provided by NCBI. Transcript abundances were estimated using Cufflinks (version 2.1.1). Cufflinks was run with the same reference annotation with TopHat2 to generate FPKM (fragments per kilobase per million mapped reads) values for known gene models.

Skin biopsies were mechanically crushed with 7 mm metallic beads in the TissueLyser equipment (Qiagen), and total RNA harvested using the RNeasy Fibrous Tissue kit (Qiagen). Sequencing libraries were prepared individually using TruSeq RNA Sample Preparation kit v2 guide (Illumina, San Diego, CA) with standard protocols, clustered using TruSeq PE Cluster Kit v3-cBot-HS kit (Illumina, San Diego, CA) and sequenced in a paired-end mode (2 × 100 bp) on a HiSeq1500 equipment (Illumina, San Diego, CA). Reads passing the CASAVA software (Illumina, San Diego, CA) quality controls were used for data analysis.