Figure 5

miR-126 shuttled by exosomes modulates VCAM1 expression in HUVECs. a: Real Time PCR analysis showed a time dependent modulation of VCAM1 mRNA expression after the addition of 10 (10 μg/ml), 20 (20 μg/ml) and 50 μg/ml (50 μg/ml) exosomes to endothelial cells. b: Real Time PCR analysis of VCAM1 mRNA expression levels in HUVECs transfected with miR-126 inhibitor (2-O-Me-miR-126) compared with untrasfected HUVECs (Control). c: Real Time PCR analysis of VCAM1 expression levels in HUVECs transfected with miR-126 mimic compared with untrasfected HUVECs (Control).d: Histogram shows the MFI (mean fluorescence intensity) of surface expression of VCAM1 in HUVECs after 24 hours of treatment with: low serum medium (Control), 20 μg/ml of exosomes (Exo 20 μg/ml). Surface expression of VCAM1 was evaluated, with FACS analysis, in HUVECs transfected with miR-126 inhibitor (2-Ome-miR-126) and treated with 20 μg/ml of exosomes (2-Ome-miR-126 + Exo 20 μg/ml). As a negative control, miScript Inhibitor Negative Control (scramble) was used. Surface expression of VCAM1 was also evaluated in HUVECs transfected with miR-126 mimic (miR-126 mimic) and treated with 20 μg/ml of exosomes (miR-126 mimic + Exo 20 μg/ml). Values are the mean ± SD of 3 independent experiments *p ≤ 0.05 **p ≤ 0.01.