Additional file 9: of Tumor stem-like cell-derived exosomal RNAs prime neutrophils for facilitating tumorigenesis of colon cancer

Figure S7. Acquisition of an immunosuppressive phenotype in bone marrow-derived neutrophils upon administration of SDCSC-exosomes. (A) T-cell suppression assay. Left: histogram showing proliferation of CD4-T cells treated with PBS or CD3/CD28 beads. Data represent the mean ± S.D. *P<.05. Right: histogram showing the proliferation of CD4-T cells treated with CD3/CD28 beads under indicated conditions. PBS, PBS-treated; Beads, CD3/CD28 bead; CM beads, CD4-T cells were treated with condition medium from PBS-treated neutrophil in the presence of CD3/CD28 beads; SDCSC Ex-CM beads, T-cells were treated with condition medium from SDCSC-exosome-treated neutrophils in the presence of CD3/CD28 beads. Data represent the mean ± S.D. *P<.05. (B) RT-qPCR under the indicated conditions. CM, CD4-T cells treated with condition medium from PBS-treated neutrophils; CM beads, CD4-T cells treated with condition medium from PBS-treated neutrophils with CD3/CD28 beads; SDCSC-Ex-CM beads, CD4-T cells treated with condition medium from CT26-SDCSC-exosome-treated neutrophil and with CD3/CD28 beads. Data represent the mean ± S.D. *P<.05. (C) GSEA results showing the association between the top 500 SDCSC-activated neutrophil signature and G-MDSC (left panel) or TAN (right panel) profiles. The expression profiles of TAN, G-MDSC and BM-Neu were from GSE43254. ES, enrichment score; NES, nominal enrichment score; FDR, false discovery rate; TAN, tumor-infiltrated neutrophil; G-MDSCs, granulocytic myeloid-derived suppressor cells; BM-Neu, bone marrow-derived neutrophils. (D) The relative viability of CD3/CD28-activated splenocytes. Splenocytes were treated with 20 μg/ml of CT26-SDCSC-exosomes or PBS in the presence of CD3/CD28 beads for 72 h. Data represent the mean ± S.D. (PDF 490 kb