Fig. 2

hEVs affect migration and invasion ability of NB cells. All experiments are performed on SK-N-AS and SK-N-DZ cells treated with EVs from SK-N-AS and SK-N-DZ cells cultured in normoxic, hypoxic and reoxygenation conditions. A. Representative images of wound healing assay; images were acquired at time zero (t0), 24 (not shown) and 48 h after treatment. Scale bar, 100 µm. B. Wound closure areas and C. cell velocity (μm/h). *p < 0.05 and ***p < 0.001 vs normoxia. D, E. Quantification of cells migrated toward the Matrigel layer (average of 5 picture fields at 10 × magnification). All values are mean ± SEM from at least 3 independent experiments. F. Representative images of colony forming assay and quantitative. Values are expressed as median ± SEM from at least 3 independent experiments. *p < 0.01 and **p < 0.01 vs normoxia. G. Relative mRNA expression of the genes involved in EMT and invasiveness (TWIST, SNAIL1, VIMENTIN and CDH1) were measured by qPCR in SK-N-AS cells after treatment with EVs from SK-N-AS cells cultured in normoxia (grey box), hypoxia (red box), and reoxygenation (blue box) conditions. Values from at least n = 3 independent experiments. *p < 0.05, **p < 0.01 and ***p < 0.001 vs normoxia. All quantifications were performed with Image J