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YOUR DATA: ,,, Direct Real-time RT-PCR on Sympathetic Neuron Lysates,,, ,,, Silencing of Focal Adhesion Kinases with Silencer Validated siRNAs

phenotypes of glioma cells. In an effort to define the contribution of each of these kinases, Dr. Joseph Loftus employed Silencer Validated siRNAs targeting PTK2 and PTK2B to knock down the expression of each of these kinases.

As seen in Figure 2, the expression of PTK2B was significantly reduced following transfection of cells with the PTK2B validated siRNA, but PTK2B expression was unchanged relative to the control in cells transfected with the PTK2 validated siRNA. Similarly, PTK2 expression was significantly reduced following transfection of cells with the PTK2 validated siRNA but unchanged following transfection with the PTK2B validated siRNA. Successful suppression of PTK2 and PTK2B gene expression will allow future analysis of the role of these kinases in integrin signaling and other cellular phenotypes.

Figure 2. Specific Gene Silencing of the Related Focal Adhesion Kinases FAK and Pyk2 . HeLa cells were transfected with PTK2, PTK2B, or a control (GAPDH) validated siRNA at a final concentration of 25 nM. 48 hours after transfection, cells were harvested and protein expression levels were analyzed by Western blot with antibodies specific for Focal Adhesion Kinase, FAK (PTK2), or Proline-rich tyrosine kinase 2, Pyk2 (PTK2B).


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