title: -Seq of mus musculus: PBS
design: Fresh tumors in the groups of PBS, NRGO@LG+L and S-G@LG+L were dissected and lysed in TRIzol reagent and followed by total RNA extraction. Purified RNA was processed with poly T-mRNA capture prior to cDNA synthesis. Sequencing libraries were generated using NEBNext Ultra RNA Library Prep Kit for Illumina (NEB, USA, Catalog: E7530L) following manufacturers recommendations. The qualified libraries were pooled and sequenced on Illumina platforms with PE150 strategy in Novogene Bioinformatics Technology Co., Ltd (Beijing, China). The raw RNA-Seq data were processed using Trimmomatic (v0.38) to remove low-quality reads and potential adaptor contaminations with the default parameter. FastQC (v0.11.8) was used to visualize the quality of processed RNA-Seq data. Then the trimmed reads were aligned to the mouse reference genome assembly build GRCm38 (mm10) using hisat2 (v2.0.5) with default parameters. The mapped reads were annotated to gencode gene exons (gencode.vM28.annotation_tran.gtf) and counted for each gene by featureCounts (v1.6.0).