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Figure 3


Fig. 3. PKC{alpha}-deficient REF cells disassemble stress fibres and focal adhesions under serum-free conditions. (A) REF cells were transfected with control siRNA oligonucleotide (–ve) or siRNA oligonucleotides 2 and 4 at the indicated concentrations. 48 hours after transfection, cells were lysed and immunoblotted for PKC{alpha} and actin. The amount of PKC{alpha} relative to the control, normalized to the actin content, is indicated below each lane. (B) REF cells were transfected with control (–ve) or siRNA duplex 4 (oligo 4) and serum starved for 16 hours. Cells were subsequently left untreated (–) or treated with LPA for 30 minutes (+LPA), fixed and stained for F-actin and {alpha}-actinin, as indicated in the Materials and Methods. Boxed areas have been enlarged to highlight the filamentous and focal adhesion localization of {alpha}-actinin. Identical results were also observed with oligo 2 (not shown). Bar, 50 µm.





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