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. 2015 Apr 17;10(4):e0123987. doi: 10.1371/journal.pone.0123987

Fig 5. HRMECs treated with SUZ12 siRNA demonstrate increased miR-200b expression, decreased VEGF expression and decreased endothelial branching.

Fig 5

A,B: Gene knockdown of EZH2 and SUZ12 was confirmed by qPCR. C,D: In HRMECs transfected with EZH2 siRNA in HG, miR-200b and VEGF were not significantly different from HG+control siRNA but decreased compared to NG+control siRNA. In HRMECs transfected with SUZ12 siRNA in HG, miR-200b was significantly increased with decreased levels of VEGF compared to HRMECs transfected with control siRNA, with levels similar to NG+control siRNA. E,F: Tube formation assay to measure endothelial branching. HRMECs transfected with HG+control siRNA demonstrated significantly increased branching compared to NG+control siRNA. Transfection of EZH2 siRNA did not reduce endothelial branching significantly compared to HG+control siRNA. However, transfection of SUZ12 siRNA significantly reduced endothelial branching compared to HG+control siRNA. [NG+control siRNA = 5mM D-glucose + 100nM control siRNA, HG+control siRNA = 25mM D-glucose + 100nM control siRNA, HG+EZH2 siRNA = 25mM D-glucose + 100nM EZH2 siRNA, HG+SUZ12 siRNA = 25mM + 100nM SUZ12 siRNA; identical letters represent groups that are not significantly different; p < 0.05; n = 6; data expressed as mean ± SEM, normalized to U6 or β-actin and expressed as a fold change of NG+control siRNA].