Fig. 7. Mon2p associates with large protein complexes and forms oligomers. (A) Native lysate from JEY001 cells (Mon2p-HA; 100 OD₆₀₀ units) was cleared by ultracentrifugation at 100,000 g for 1 hour and subjected to gel filtration chromatography. Fractions were analyzed by SDS-PAGE and western blotting using a monoclonal anti-HA antibody. All fractions containing Mon2p-HA are shown and approximate complex size (based on a prior run of sizing standards) is indicated. (B) Mon2p-myc coimmunoprecipitates with Mon2p-HA. Detergent-solubilized native lysates from cells (50 OD₆₀₀ units) expressing Mon2p-myc alone (JEY003) or with Mon2p-HA (JEY026) were cleared at 16,000 g and subjected to immunoprecipitation using anti-HA antibody. Purified proteins were subjected to SDS-PAGE and western blotting analysis with anti-myc and anti-HA antibodies.