EpCAM knockdown using EpApt-siEp construct in WERI-Rb1 and MCF7 inhibits cell proliferation.

<p><b>A.</b> The EpCAM mRNA levels were detected from the total RNA of control, siEp and EpApt-siEp treated MCF7 cells by northern blotting. The total RNA was electrophoresed in formaldehyde agarose gel electrophoresis, blotted and developed by chemiluminescence based method. EpCAM targeting siRNA was used for synthesizing probe. On its right, the densitometry analysis of the bands were performed using imageJ software and plotted as graph with % mRNA expression against the 28s rRNA. <b>B</b>. The EpCAM mRNA levels were quantified by SYBR green based qPCR from the cDNA of control, siEp and EpApt-siEp treated WERI-Rb1 and MCF7 cells. <b>C</b>. Western blotting was performed on the siEp transfected and EpApt-siEp treated WERI-Rb1 and MCF7 cells for the EpCAM and b-tubulin. EpCAM targeting siRNA was used for synthesizing probe. <b>D</b>. The densitometry analysis of the western blotting bands was performed using imageJ software and plotted as graph with % protein (EpCAM) expression normalized to β-tubulin. <b>E</b>. The percentage cell proliferation was quantified by performing MTT assay on the control, siEp transfected, EpApt-siEp, EpApt and ScrApt treated WERI-Rb1 and MCF7 cells. The graph shows the % cell proliferation normalized to control cells. Data represents mean ± SD. Experiments were repeated 3 times independently with similar results. **P value of 0.01–0.001; *P value of 0.05–0.01.