Figure 8.

Time-course analyses of siRNA cellular localization mediated by RD3AD in comparison to 599

Confocal fluorescence microscopy analyses of CAL 27 cancer cells 0.5, 1, 2, and 4 h post-treatment with DY547-siCIP2A (red) in complex with 599 or RD3AD at a 50:1 Peptide:siRNA molar (12.5 N/P) ratio. Alexa Fluor 488 phalloidin was used to stain the F-actin (green) in filopodia, as well as the cytoplasm of cells, so as to help demarcate the cell bodies. Nuclei (blue) were counterstained with DAPI. The upper-half panels represent merged images of the red and blue fluorophore signals only, while the lower-half panels represent all three (red, blue, and green) fluorophore signals combined. Cross-sectional views comprising z sections along the x and y planes are also presented within each panel. Yellow fluorescence reveals the potential colocalization between siRNA and F-actin. Scale bar: 70 μm.