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Fig. 5 | Journal of Hematology & Oncology

Fig. 5

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

Fig. 5

Systemic biology analysis identifies expression of exosomal RNAs-induced interleukin-1β is required for neutrophil survival. a Viability of neutrophils treated with different condition medium of educated-neutrophils. PBS-CM, conditional medium from PBS-treated neutrophil; SDCSC-Ex-CM, condition medium from SDCSC exosome-treated neutrophils. ***P < .001. b Validation of IL-1β expression using RT-qPCR (left panel) and ELISA (right panel) under indicated conditions. **P < .01. c A histogram showing neutrophil viability under indicated conditions. IgG, normal IgG control (10 μg/ml); Il-1b nAb, neutralizing IL-1β antibody. *P < .05, **P < .01. d Representative images for showing size distribution of exosomal RNA by Agilent Bioanalyzer. nt, nucleotide. e Pie charts showing exosomal RNA context of parental cells and SDCSCs. miRNA, microRNA; piRNA, Piwi-interacting RNA; snoRNA, small nucleolar RNA; snRNA, small nuclear RNA; srpRNA, signal recognition particle RNA; scRNA, small cytoplasmic RNA; tRNA, transfer RNA; LINE, long interspersed nuclear elements; SINE, short interspersed nuclear elements; LTR, long terminal repeat. f RT-qPCR examining Il1b expression in neutrophils upon transfection. Cellular and exosomal RNAs were extracted from CT26-SDCSCs. CIP, calf intestinal phosphatase. *P < .05, **P < .01. g RT-qPCR examining Il1b expression in neutrophils. Act D, actinomycin D (0.3 μg/ml). ***P < .001. h Flow cytometry results for showing expression of IκBα. i Left: Representative images illustrating nuclear p65 expression. Right: a histogram for showing percentage of nuclear p65(+) cells. ***P < .001. j RT-qPCR showing Il1b expression in neutrophils upon blocking NFκB pathway. Exosomal RNA was extracted from CT26-SDCSCs. Parthenolide, a NFκB inhibitor (Par, 0.3 μM). Cells were transfected with 100 ng of exosomal RNAs for 6 h followed by parthenolide or DMSO treatment for a total of 24 h. *P < .05, **P < .01

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