RNA-Seq of skin biopsy
RNA was extracted from the samples following the RNeasy Fibrous Tissue Mini kit protocol (Qiagen) with a DNase I (Qiagen) treatment. Quality assessment was carried out using the NanoDrop 2000 (Thermo Fisher), visualized through gel electrophoresis and the TapeStation system with RNA Screen Tape (Agilent Technologies), and concentrations were quantified using the Qubit 2.0 fluorometer (Invitrogen). The cDNA library was prepared following the TruSeq Stranded mRNA Library Prep kit protocol (Illumina, Inc). The cDNA quality was validated through the Bioanalyzer 2100 instument. cDNA library quantification was performed through qPCR, following the Sequencing Library qPCR Quantification kit instructions (Illumina Inc.). The sample was diluted to a concentration of 10 pM and clustered on cBot. Sequencing was carried out on the Illumina HiSeq 2000 system, with paired-ended reads PE 2 x 100 bp