RNA sequencing analysis of CAF treated by colon cancer cell-derived exosomes

In this study, we analyzed differentially expressed genes by obtaining gene expression values through transcriptome sequencing of Homo sapiens, and performed functional classification and gene annotation for significant genes based on gene ontology and pathway information. After the pre-processed trimmed reads were mapped to a known reference genome using the HISAT2 program, transcript assembly was performed through the StringTie program. As a result, expression profile values were obtained for each sample for the known transcript, and read count, based on transcript/gene Fragment per Kilobase of transcript per Million mapped reads (FPKM), Transcripts per Kilobase (TPM) Million) values have been summarized. This value was subjected to DEG (Differentially Expressed Genes) analysis using edgeR for comparison combinations (HT-29_Exo-CAF vs. CTL_PBS-CAF, LoVo_Exo-CAF vs. CTL_PBS-CAF, and SW480_Exo-CAF vs. CTL_PBS-CAF), and genes that satisfies the condition |fc|>=2 & exactTest raw p-value<0.05 in at least one comparison combination Dogs were extracted. Transcriptome resequencing data was used to compare expression profiles between comparable samples. Gene Ontology Enrichment analysis was performed using the g:Profiler tool (https://biit.cs.ut.ee/gprofiler/) for a list of genes with significant expression level differences. GO_stat is the result of organizing the associated gene and test stat based on term_id. GO_genes is the result of arranging the associated term_id and DEG analysis result stat based on the gene.