MiR-27a Targets VE-cadherin. (A) Expression of VE-cadherin in control or miR-27a-mimic transfected cells after 48 hours. β-actin was used as a loading control. (B) The normalized expression of the mean of 5 independent human umbilical vein entothelial cell (HUVEC) lines ± standard error of the mean (SEM). *P < .05 cf control. (C) The level of surface VE-cadherin as assessed by flow cytometry. The solid line represents the control mimic; the dashed line represents the miR-27a mimic. This is the result of one experiment, similar to three performed. (D) The level of VE-cadherin mRNA expression 24 hours posttransfection with miR-27a mimic. Results normalized to β-actin are the mean of quadruplicate quantitative reverse-transcription polymerase chain reaction (qRT-PCR) reactions ± SEM from 5 independent HUVEC lines. *P < .05 cf control. (E) VE-cadherin expression from HUVEC 48 hours post transfection with control or LNA-27–transfected cells. (F) The mean of 3 independent HUVEC lines ± SEM is shown. ***P < .001 cf control. (G) The level of surface VE-cadherin as assessed by flow cytometry. The solid line represents the control LNA; the broken dashed line represents LNA-27. This is the result of one experiment similar to two performed. (H) Luciferase constructs containing the 3′UTR of VE-cadherin containing the putative miR-27a binding site (Wt, black bars) or a mutated miR-27a binding site (Mut, white bars), together with control or miR-27a mimic. Results represent the mean of triplicate transfections ± SEM from 4 independent experiments. *P = .01, Wt + miR-27a vs Mut + miR-27a. **P = .00001, Wt + control vs Wt + miR-27a.