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Fig. 4 | Molecular Cancer

Fig. 4

From: Exosome-mediated microRNA signaling from breast cancer cells is altered by the anti-angiogenesis agent docosahexaenoic acid (DHA)

Fig. 4

DHA-induced exosome microRNA transfer from MCF7 cells to endothelial cells alters microRNA expression, endothelial tube formation, and target mRNA expression. a EA.hy926 cells (2.5×104) were plated in a 24-well plate and MCF7 cells (2×104) were plated in a transwell insert (0.4 μM) and co-cultured for 24 h. MCF7 cells were treated with 100 μM DHA for 24 h, RNA was extracted and qRT-PCR was performed. The fold change in expression levels was calculated using the ΔΔCt method. Data in bar graphs represent mean ± SEM (n = 3). EA.hy926 cells were transfected with a scramble control, let-7a, miR-21, miR-23b, miR-27b, or miR-320b mimics. After 48 h EA.hy926 cells (1×104) were plated on ECMatrix in a 96-well plate and incubated at 37 °C for 16–18 h. Branch points were counted (b) and representative images (c) were taken and under a light microscope (20×). Shown are the combined triplicate results from three independent experiments. Data in bar graphs represent mean ± SEM (n = 9). *, p < 0.001 using Student’s t test. d EA.hy926 cells (2.5×104) were seeded on a 24-well plate and MCF7 cells (2×104) were cultured overnight in a transwell insert (0.4 μm). MCF7 cells in the upper chamber were treated with DHA for 24 h. RNA was extracted from the EA.hy926 cells in the lower chamber and target gene expression was measured by qRT-PCR. EA.hy926 cells were transfected with a scramble control, miR-23b, or miR-320b mimic. The previously validated mRNA targets (PLAU and AMOTL1) for miR-23b (e) and (NRP1 and ETS2) miR-320b (f) were measured by qRT-PCR 48 h post-transfection. The fold change in expression levels was calculated using the ΔΔCt method. Data in bar graphs represent mean ± SEM (n = 3). *, p < 0.05 using Student’s t test

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