Figure 3.

Protease protection assay. FKBP65 is entirely intravesicular. The 200,000g microsomal pellet from NIH3T3 cells was treated with buffer alone (control), 100 μg/ml proteinase K alone, or 100 μg/ml proteinase K + 1% Triton X-100. After 30 min at room temperature, excess PMSF was added to each sample to inhibit the enzyme, and the membranes in all samples were solubilized with Triton X-100. Proteins were acetone precipitated and Western blotted. CNX-CT = calnexin C-terminal antibody.