Figure 2.

Syt VII colocalizes with dense lysosomes on subcellular fractionation. a, Western blot of NRK fibroblasts, L6E9 myoblasts, and rat brain extracts (5 μg/lane), and recombinant Syt VII (rSyt VII) with anti-Syt VII antibodies in the presence of the scrambled control peptide (left), or in the presence of the NH₂-terminal Syt VII peptide (center). Right, Western blot with antibodies to Syt I (MC17) of the same extracts, and recombinant Syt I (rSyt I). b, Percoll density gradient fractionation of NRK cells. Top, detection of β-hexosaminidase, mannosidase II, and transferrin–HRP on gradient fractions collected from the bottom (activities assayed using 4-methyl-umbellyferyl-N-acetyl-β-d-glucosaminide, 4-methylumbelliferyl a-D-mannopyranoside, and 3,3′, 5,5′-tetramethylbenzidine as substrates, respectively). Bottom, Western blot detection of Syt VII, cathepsin L, rab 7, EEA1, and calnexin on gradient fractions.