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. 2015 Nov 12;5:16437. doi: 10.1038/srep16437

Figure 7. Knockdown of A20 induces the apoptosis of MDSCs through JNK pathway.

Figure 7

(a) Isolation of MDSCs from tumor tissue was confirmed by flow cytometry. Total number of 30000 cells was analyzed. Numbers illustrated indicate the percentage of the cells in total cells (n = 5). (b) mRNA levels of A20 from the isolated MDSCs from spleen and E.G7 tumor were measured (n = 3). (c) Fluorescence immunostaining of Gr1 and A20 positive cells in MDSCs isolated from tumor tissue. Scale bar 50 μm. (d) Si-A20 treatment induced the apoptosis of MDSC in vitro. Gr1+CD11b+ cells were isolated from tumors in E.G7 tumor-bearing mice and transfected with si-RNA. Apoptosis of MDSCs were confirmed by flow cytometry. Total number of 30000 cells was analyzed. Numbers illustrated indicate the percentage of the cells in total cells. MDSCs were cultured with a mixed medium (TCCM: complete RPMI 1640 = 1:1) with GM-CSF (10 ng/ml). Percentages of the cells in the regions were illustrated. (e) Statistic analysis of (d). (f) Si-A20 induced the apoptosis of MDSCs through JNK pathway. p-JNK, activated caspase-3 and activated caspase-8 were analyzed by western blotting after treatment of MDSCs with si-A20 and TNF-α. Data were representative of two independent experiments. Data represent means ± SD. *p < 0.05 (ANOVA test, Student’s t-test).