title: RNA-Seq of B16 cells stimulated with IL17A 100ng/mL 24h
design: B16 and EL4 cells cultured under serum deprivation conditions with IL-17A (100ng/mL) or medium alone for 24h, the medium was removed, harvested, and lysed with TCL buffer (Qiagen) with 1%  -mercaptoethanol and stored at -80 C until further analysis. RNA was isolated using a single cell RNA purification kit (Norgen), which included treatment with the RNase-Free DNase Set (Qiagen). RNA integrity number was assessed with an Agilent RNA 6000 pico kit (all RIN>8). Reverse transcription was performed with the SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing. Barcoded Illumina-compatible libraries were generated from 5 to 10 ng of DNA from each sample, using the Nextera XTP Repair Kit. Libraries were sequenced on an Illumina Novaseq 6000 using 100 bp paired-end mode, 20 million reads per sample.