RNA-seq of xenograft tumor in control group and C26 group

We performed RNA-sequencing (RNA-seq) with tumor tissues of mice MC38 xenograft models treated with C26. Tumor tissues of mice MC38 xenograft models were utilized for total RNA extraction employing the RNeasy plus mini kit (Qiagen, #74134). Post-extraction, the RNA samples underwent evaluation of purity and concentration via 4200 TapeStation analysis. Subsequently, RNA-Seq samples were pooled and subjected to sequencing on NextSeq2000 using Illumina TruSeq Stranded mRNA Library Prep, employing paired-end sequencing services provided by the National Cancer Institute Core Sequencing Facility. Reads from the samples underwent adapter and low-quality base trimming using Cutadapt prior to alignment against the reference genome (mm10) and annotated transcripts using STAR. Mapping statistics were computed using Picard software, while library complexity was assessed based on unique fragments in the mapped reads using Picard’s MarkDuplicate utility. Furthermore, gene expression quantification analysis was conducted for all samples using STAR/RSEM tools. Raw counts were subsequently uploaded for gene expression and KEGG pathway analysis with online NetworkAnalyst platform (https://www.networkanalyst.ca).