HUVECs were treated with plasmids and 5.5 or 30 mM glucose. when the cells were 90-95% confluent, a sterile 200 μl pipette tip was used to draw straight lines in the middle of the well, and serum-free ECM medium was used to wash the cells twice, and this was then replaced with ECM medium containing 2% FBS, 1% PS and 1% EGF, as previously described (25). Images were collected (Canon EOS 600d) at this time as a 0 h control. The culture was continued for a further 24 h at 37°C. The image data were collected after 24 h.

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