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Figure 2. Time-course for the digestion of rpAMBN by rpMMP-20. (A) 4–12% NuPage gels stained with CBB (left) and silver (right). (B) Western blots of a similar time-course immunostained with the AMBN N-terminal (AMBN-63) and C-terminal (AMBN-381) anti-peptide antibodies. The lanes are marked according to the times (in hrs) of the digestions. An asterisk marks negative controls with no rpMMP-20 added. The rpAMBN incubated without rpMMP-20 was stable and was undigested for the duration of the time-course, indicating that rpMMP-20 was solely responsible for the AMBN cleavages in the experimental conditions. Western blot analyses with region-specific anti-peptide antibodies showed that the smeared band at 48 kDa did not contain the AMBN N-terminus, while the smaller bands at 16, 18, and 19 kDa did.
