FIG. 3.

Western blot analysis of tagged Mig1, Mig2, and Mig3. Strains YM6843 (MIG1-6HA), YM6833 (MIG2-6HA), and YM6835 (MIG3-6HA) were pregrown in 5% glycerol plus 0.5% galactose (no glu) and inoculated into the same medium and into a medium containing 5% glycerol plus 0.05% glucose (low glu) or 4% glucose (high glu) at an OD₆₀₀ of 0.1 to 0.2. Protein extracts were prepared after the cultures reached an OD₆₀₀ of approximately 1.0. For the MIG1-6HA strain, each lane was loaded with extract in a volume equivalent to a cell OD₆₀₀ of 0.25. For MIG2-6HA, cells with an OD₆₀₀ of 0.5 were loaded; for MIG3-6HA, cells with an OD₆₀₀ of 1.0 were loaded. Proteins were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (4 to 15% polyacrylamide gradient) and detected by immunoblotting. Loading of the lanes with expected amounts of protein was subsequently confirmed by staining the membrane with Ponceau Red (data not shown).