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Fig. 7. Furin is excluded from detergent-resistant membranes. (A) Untransfected (NT) and furin-GFP-transfected (FUR) A375 cells were labeled with [³⁵S]methionine for 2 hours and lysed with TNE buffer containing 1% Triton X-100 on ice. Soluble (S) and insoluble (P) fractions were separated by ultracentrifugation, immunoprecipitated with a polyclonal anti-GFP antibody and analyzed by SDS-PAGE/autoradiography. (B) A375 cells transfected with MT1-MMP (MT1-MMP) or with MT1-MMP and furin-GFP (MT1+FUR) were labeled with [³⁵S]methionine for 2 hours and lysed with TNE containing 1% Triton X-100 on ice. Soluble fractions were collected after ultracentrifugation, immunoprecipitated with polyclonal anti-MT1-MMP antiserum MMR2 and analyzed by SDS-PAGE/autoradiography. The processing ratio of MT1-MMP (MT1-MMP/MT1-MMP+proMT1-MMP) was determined with the Fuji BAS software and plotted on the reported chart.

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