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Fig. 2 | Stem Cell Research & Therapy

Fig. 2

From: Non-stem bladder cancer cell-derived extracellular vesicles promote cancer stem cell survival in response to chemotherapy

Fig. 2

NSCC-derived EVs affect CSC properties. a EVs were purified from MB49 F1 and S2 cells, and their purity (10 µg of total EVs proteins) was confirmed by Western blotting for exosome markers (TSG101 and CD9) and ER-resident proteins (Bip and PERK) to exclude potential cytoplasmic contamination. b Determination of EV secretion rate in MB49 F1 and S2 cells by NTA. c The effect of EVs on MB49 S2 cell sphere formation was measured by seeding MB49 S2 cells (1000 cells/mL) in CSC culture medium and 10 µg/ml EVs were used to treat S2 CSCs for 21 days. Images of spheroid formation on days 1, 7, and 21 are shown (50x). Spheroid numbers were counted and the resultant quantitative data were plotted. d The expression of CSC markers in the S2 cells shown in C. PBS treatment was used as a normalization control, and relative expression was calculated via the 2−ΔΔCt method. e F1-EVs promote MB49 S2 cell migration and invasion. S2 cells maintained in CSC medium were treated with F1-EVs, S2-EVs, or PBS for 10 days, cells were subjected to migration and invasion assays using 10% FBS as a chemoattractant. For migration assay, cells were incubated for 48–72 h. Experiments were repeated independently at least three times. Data were compared via Student’s t-tests or one-way ANOVAs with Tukey’s multiple comparisons test, as appropriate. *P < 0.05; **P < 0.01; ***P < 0.001

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