Fig. 3

RNA-seq transcriptome analysis identified the differentially expressed genes in tumor treated with CCL11-E6E7, E6E7 or vehicle. After the grafted TC-1 tumor is palpable, one dose of 25 µg CCL11-E6E7, E6E7, or an empty vector was intramuscular injection followed by electroporation. The tumor was removed and subjected to RNA-seq analysis on the fourteenth day. A Volcano plot was used to visualize the DEGs in tumor tissue immunized with CCL11-E6E7 compared to E6E7. The red dots represent significantly up-regulated genes and the blue dots represent significantly down-regulated genes as calculated by |log2 FC|> 0 and FDR < 0.05. B The GO enrichment analysis was performed on genes that were significantly up-regulated by CCL11-E6E7 vaccination. Enriched pathways were determined with a significance cutoff of p-value < 0.05 (Fisher’s exact test) and a requirement for at least 40% up-regulated genes in the pathway. C The upregulated DEGs extracted from CCL11-E6E7 vs E6E7 that are positive associated with M1 macrophages and stem-like T cell activation in the tumor tissue were used to perform hierarchical clustering. D TCR complementarity-determining region 3 (CDR3) length distribution patterns among TCRA and TCRB reads in each group. The x-axis represents the CDR3 length in nucleotide (nt) and the y-axis represents the number of each length group in all the identified CDR3 sequences. E, F The T cell receptor (TCR) clonotypes showed increased abundance and expansion upon CCL11-E6E7 treatment compared to E6E7 treatment. The TCRA and TCRB sequence analysis was used to quantify the number of TCR clonotypes detected in each treatment group (n = 3/group). The chao1 indexes were calculated as a measurement for TCRA and TCRB clonal diversity. The p-value was measured by Student's t-test