Bulk RNA-Seq of actinic keratoses

RNA-Seq was performed on 8 actinic keratoses (AK) to identify differentially expressed genes in AK vs normal skin and cutaneous squamous cell carcinoma Total RNA was then isolated from actinic keratoses using QIAshredder (Qiagen) and Trizol (ThermoFisher) followed by DNA removal with the TURBO DNA-free ki t (Ambion) according to the manufacturer’s instructions. RNA integrity was verified using an Agilent 2100 Bioanalyzer. RNA-Seq libraries were prepared with the mRNA Seq Sample Prep K it (Illumina, Inc.) as recommended by the manufacturer. 101 bp paired-end sequencing reads were obtained using the Illumina HiSeq platform.