Fig. 2. Time-course analysis of MT1-MMP processing in A375 melanoma cells and association with different membrane fractions. Untransfected (NT) and MT1-MMP-transfected A375 cells were pulse-labeled with [³⁵S]methionine for 5 minutes and, after the indicated chase times, incubated with TNE containing 1% Triton X-100 on ice. Soluble (S) and insoluble (P) fractions were separated by ultracentrifugation, immunoprecipitated with polyclonal anti-MT1-MMP antiserum MMR2, subjected to SDS-PAGE, transferred to nitrocellulose membrane and visualized by autoradiography. The chart shows the distribution in time of immature and mature MT1-MMP and association with different membrane fractions as determined with the Fuji BAS software.