RNA-seq of  MB49 tumor aPD-1 resistant
RNA was  extracted using RNeasy mini kit (Qiagen Cat. No. 74104). Quality of samples  was checked by Bioanalyzer 2100 (Agilent) and RNA was quantified by Nanodrop.  After, 5 g of RNA was enriched by NextFlex Rapid Directional mRNA-Seq  (Bioo-Scientific) kit. Quality of samples were checked again by Bioanalyzer  2100 (Agilent) and quantified by qPCR. "Samples were put on Flow Cell  High Output. Amplification and sequencing were performed with Illumina  NextSeq500: run Paired-end (2*75nt) for all MB49 tumor and run Single Read 75  bp for MC38."