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Figure 2


Fig. 2. Overexpression of dominant-negative PKC{alpha} results in stress fibre disassembly under serum-free conditions. (A) REF cells were transfected with a plasmid encoding dnPKC{alpha}-GFP fusion protein. 24 hours after transfection, cells were either grown with (+FCS) or without (–FCS) serum for a further 16 hours. Some serum-starved cultures were treated for 30 minutes with LPA (400 ng/ml), fixed and stained for F-actin. (B) REF cells were transfected with wtPKC{alpha}-GFP and treated as described above. Transfected cells retain stress fibres in the presence or absence of serum. Bar, 50 µm. (C) Quantification of the percentage of cells expressing the PKC{alpha}-GFP constructs undergoing stress fibre disassembly. Black bars, dnPKC{alpha}-GFP; white bars, wtPKC{alpha}-GFP. LPA stimulation was not performed on cells transfected with the wt construct and, therefore, quantitative results are not shown. Results were obtained from three independent experiments (± s.d.).