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Fig. 1. Treatment of REF52 cells with siRNAs for FAK results in loss of FAK expression and inhibition of focal adhesion protein phosphorylation. (A) REF52 cells were untreated, or treated with either luciferase or FAK siRNA, and equal amounts of lysates were analyzed by western blot 72 hours later for FAK expression (a). The blot was stripped and reprobed with an anti-phosphotyrosine antibody (P-Tyr, b), anti-phospho-FAK 397 (c), anti-phospho paxillin Y31 (d) and anti-phospho-paxillin Y118 (e). The blot was also stripped and re-probed for total paxillin (f) and p130Cas levels (g) as loading controls. Cells were either kept in suspension (S) or allowed to adhere to fibronectin (FN) for the indicated times. (B) Cells treated with either the luciferase siRNA (a,c) or FAK siRNA (b,d) were allowed to adhere to fibronectin-coated coverslips and immunofluorescently stained for either FAK (a,b) or actin (c,d). The arrows (a) denote focal adhesions.
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