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Fig. 9. Cre-mediated deletion of FAK in primary fibroblasts results in impaired leading edge formation. (A) PCR analysis of wild-type (lanes 1-2), homozygous floxed (lanes 3-4) or heterozygous floxed (lanes 5-6) fibroblasts. Lanes 1, 3 and 5 show the PCR products from DNA of cells that were not exposed to the Cre-containing adenovirus; lanes 2, 4 and 6 show cells the products from cells infected with adeno-Cre. Arrow points to the band corresponding to the expected PCR product from the intact FAK/Lox sequence; arrowhead indicates the PCR product from the recombined sequence following Cre-mediated excision. (B) Western blot of cellular lysates from wild-type, floxed and heterogeneous WT/Flox fibroblasts. Arrow indicates the band corresponding to FAK. The blot was stripped and probed for ERK as a loading control. Monolayers of either wild-type (C) or Floxed (D) fibroblasts were wounded and analyzed by video time-lapse microscopy. The white outlines denote the plasma membranes of the cells that started from the edge of the wound at the bottom of the photograph. See Movies 5 and 6 in supplementary material.





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