Inhibition of gene expression by siRNA is mostly mediated by the use of a transfection reagent,
delivering the siRNA into the cytoplasm of the cell. Most protocols show optimized data using
25 to 100 pmol of siRNA per 6-well. (Final concentration of siRNA is 25 to 100nM)

We compared several siRNA transfection reagents by evaluating the expression inhibition of the
epithelial-glycoprotein EpCAM on SKOV-3 tumor cells.

10,000 cells were seeded in a 96 well plate and transfected with 3 pmol siRNA
(which results in a final concentration of siRNA of 15nM).

Cells were treated with an EpCAM specific siRNA and incubated for 72 h. Knock-down efficiencies
were determined by measuring the EpCAM expression level after incubation with the EpCAM specific Antibody
MOC-31 labeled with Alexa-488. Results were measured as relative fluorescence units (RFU) on a
fluorometer and percentages remaining EpCAM expression were normalized to a no-siRNA control
(set 100%, not shown).

The results clearly show that SAINT-RED mediated delivery of siRNA generates the highest
inhibition by far compared to the competitors using only 3 pmol of siRNA.
SAINT-REDs knock-down efficiency is constant from only 1 pmol to the highest dose tested, 5 pmol
(data not shown).