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Fig. 11. Isolation of disulphide-bonded trans-dimers. Magnetic beads coated with N-cadherin-Fc bearing the mutation D27C were allowed to stick to K562 cells expressing wild-type N-cadherin or the mutants D1C, R25C or D27C. After formation of disulphide bonds, the trans-dimers attached to the beads were isolated from cis-dimers and non-involved cell surface N-cadherin. The trans-dimers were detected by immunoblotting for cellular N-cadherin cytoplasmic domain. The upper panel shows that trans-dimers formed only with the combination D27C beads adhering to D1C cells. The main band at about 300 kDa represents N-cadherin-Fc (dimerised by the Fc hinge region) disulphide-bonded to one cellular cadherin molecule. Higher order assemblies are also seen. The right-hand panel shows total cellular cadherin for comparison. The lower panel shows a gel loading control.





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