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Figure 5


Fig. 5. Loss of PKC{epsilon} results in a reduced ability of fibroblasts to contract and adhere to matrix. (A) Contraction of a collagen gel matrix: FPCL analysis. The effect of loss of PKC{epsilon} expression on contractile force generated by fibroblasts in a fixed, tethered floating collagen gel lattice was investigated using a Culture Force Monitor (Eastwood et al., 1994). Force generated by PKC{epsilon}+/+ and PKC{epsilon}-/- fibroblasts was assessed over 24 hours. Traces are the result of using 5 ml of collagen gel at a density of 1 million cells per ml of collagen gel in 2% serum. A representative trace is shown (n=3). The higher contractile forces generated by cells derived from PKC{epsilon}+/+ in comparison with PKC{epsilon}-/- fibroblasts. (B) Loss of PKC{epsilon} results in a reduced ability of fibroblasts to adhere to extracellular matrix. Adhesion of PKC{epsilon}+/+ and PKC{epsilon}-/- fibroblasts on fibronectin (FN) and type I collagen (col) was assessed as described in the Materials and Methods. Cells were detached from plates with EDTA, and equal numbers of cells were placed into individual wells of 96-well plates. Adherent cells were detected (Absorbance 450) 45 minutes post-adhesion (average±s.d., n=3). *P<0.05 relative to wild-type control.





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