Figure 3.

The effect of the multiple transfections of miR-181a-2 on the ERα transcriptional activity and protein profile. The MCF7 cells were cultured for two months after multiple transfections with miR-181a-2-mimetic or scrambled-RNA construct, then the cells were transfected with the ERE plasmid containing the luciferase reporter gene under the oestrogen-responsive element (ERE) and β-galactosidase plasmid (a). The relative luciferase activity was calculated in arbitrary units as the ratio of the luciferase to the galactosidase activity. Data represent the mean value ± SD of three independent experiments. The viability of cells treated with vehicle control was set at 100%. (b) Western blot analysis. The MCF7 cells were treated as indicated above. Western blot analysis of Akt, phospho-Akt and ERα was performed in the MCF7 cell extracts. Protein loading was controlled by membrane hybridization with α-tubulin antibodies. Densitometry for immunoblotting data (right diagram) was carried out using ImageJ software; * p < 0.05 versus scrambled (scr).