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GENE SILENCING WITH PCR PRODUCTS

in c-fos Expression. siRNA Expression Cassettes encoding a negative control or c-fos-specific hairpin siRNA were transfected into HeLa cells. 72 hours post-transfection, the cells were assessed using nuclear staining with DAPI and immunofluorescence using a c-FOS antibody detected with a fluorescein labeled secondary antibody.

Figure 2. siRNA Expression Cassette-Mediated Reduction in GAPDH Expression. siRNA Expression Cassettes encoding a negative control or GAPDH-specific hairpin siRNA were transfected into HeLa cells. 72 hours post-transfection, the transfected cells and a non-transfected control sample (NT) were assessed using nuclear staining with DAPI and immunofluorescence using a GAPDH-specific antibody. The DAPI and immunofluorescence images are overlaid in the figures.

Figure 3. Variable Reduction in Target Gene Expression Using SECs with Different Promoters. siRNA Expression Cassettes featuring the mouse U6 (Mo-U6), human U6 (Hu-U6), and human H1 (Hu-H1) promoters and encoding a c-fos-specific hairpin siRNA were transfected into HeLa cells. 72 hours post-transfection, the cells were assessed using nuclear staining with DAPI and immunofluorescence using a c-FOS antibody. Non-transfected cells (NT) as well as cells transfected with an SEC expressing a negative control siRNA (scramble) demonstrate wild-type levels of c-FOS. The relative level of reduction in gene expression was quantified and is provided in the bar graph.

Figure 4. Reduction in -actin Expression Using SECs with Different Promoters. siRNA Expression Cassettes featuring the mous
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