Figure 7.

Ectopic expression of miR-133a-3p results in a decrease of pro-angiogenic and an increase in anti-angiogenic proteins in endothelial cells. (A) HUVEC were transfected with a control miR mimic (NC) or a mimic for human miR-133a-3p (3p) and were left unstimulated (−) or were stimulated with VEGF-A₁₆₅ (50 ng/ml) for 4 h (+). Expression of CCNB1, CD44 and RCAN1.4 was determined by western blot analyses of total proteins isolated from the transfected cells. Levels of Tubulin were determined in the same samples for loading control. Blot images are representative of two independent experiments. The band corresponding to Cyclin B1 is pointed out by an arrow. *, denotes unspecific binding. (B) HUVEC were transfected as in (A) and then stimulated (+) or not (−) with VEGF-A₁₆₅ (50 ng/ml) for 5 min. The expression and phosphorylation/activation status of VEGFR2 and Erk1/2 proteins was determined by western blot analysis of total proteins isolated from the transfected cells. Blot images are representative of two independent experiments. Levels of Tubulin were determined in the same samples for loading control.