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. 2014 May 16;3(3):e000968. doi: 10.1161/JAHA.114.000968

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© 2014 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley Blackwell.

This is an open access article under the terms of the Creative Commons Attribution‐NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.

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Figure 1. — AVM‐BEC‐conditioned media (AVM‐BEC‐CM) potentiates miR‐18a internalization. A, AVM‐BEC and control BEC were analyzed for intracellular miR‐18a levels using qPCR. Control BEC were used as baseline (n=3; ***P<0.001; paired t test). B, miR‐18a (40 nmol/L) in combination with AVM‐BEC‐CM (black bars) or fresh culture media (white bars) was added to AVM‐BEC and tested for intracellular miR‐18a after 5, 10, 30 minutes (n=4), 1, 2, 8 and 24 hours (n=3). AVM‐BEC‐CM enhanced miR‐18a entry up to 30 minutes (*P<0.05, **P<0.01, ***P<0.001; paired t test). C, Control BEC were treated with miR‐18a (40 nmol/L) in combination with AVM‐BEC‐CM (black bar), control BEC‐CM (gray bar) or fresh media (white bar). Intracellular miR‐18a was analyzed (qPCR) after 30 minutes incubation; AVM‐BEC‐CM potentiated miR‐18a internalization by control BEC (n=3; *P<0.05; paired t test). D, Control BEC were treated with miR‐18a (40 nmol/L) and in the presence of serial diluted AVM‐BEC‐CM (diagonal line bars), demonstrating that progressively diluted AVM‐BEC‐CM loses its ability to enhance miR‐18a internalization (n=3, *P<0.05, **P<0.01; paired t test). Dotted line represents miR‐18a uptake by control BEC in the presence of fresh media. Data are presented as mean±standard error of the mean (SEM). Each human specimen‐derived cell culture represents a unit of analysis (n). AVM indicates arteriovenous malformation; BEC, brain endothelial cells; CM, conditioned media; qPCR, quantitative real‐time polymerase chain reaction.