GSM5393816: Scar.B.RNA; Homo sapiens; RNA-Seq
RNA-seq was performed on FACS-isolated mouse fibroblast subpopulations from JUN and Control mice and human cultured KO and non-KO fibroblasts resuspended in TRIzol (10k cells/1ml). The RNA-seq data was collected from ten mice and pooled into two biological replicates. Thus, each biological replicate represents 10 wounds, as each mouse has two wounds. The human RNA-seq data was collected from 4 individual patients. RNA was isolated using the Rneasy Micro Kit (Qiagen, #74004), as per the manufacturer's instructions. ATAC-seq libraries were generated using an average of 50,000 freshly isolated fibroblasts per replicate (biological replicates for mice samples and technical replicates for human samples) as described by Buenrostro et al 2013 with the following modifications. Transposition was performed in 50 uL reaction containing in-house made 2xTD buffer, Tn5 and 1% Digitonin. Libraries were sequenced on an Illumina HiSeq 2500 machine. KO and non-KO fibroblasts from each skin type will be prepared for RNA-seq. ATAC libraries were prepared using the protocol found in ATAC-seq: A Method for Assaying Chromatin Accessibility Genome-Wide by Buenrostro et al (2015).