Figure 1

Experimental conditions to study glucose effects in regulating protein levels. A, wild-type cells expressing Lia1p-GFP were grown either in YNB-based media containing 2% glucose (I and II) or in YPKG-based media containing 0.5% glucose (III and IV) for 3d. In experiment I, 2% glucose was added directly to the existing YNB culture for 0, 2, and 4 hours. In experiment II, cells were grown in YNB-based media, pelleted and resuspended in fresh YNB with 2% glucose for 0, 2, and 4 hours. In experiment III, cells were grown in YPKG for 3d and 2% glucose was added directly to the existing YPKG media for 0, 2, and 4 hours. In experiment IV, cells were grown in YPKG for 3d, pelleted, and resuspended in YPD media for 0, 2, and 4 hours. Levels of Lia1p-GFP, Fbp1p, and Tpi1p were examined using anti-GFP, anti-Fbp1p, and anti-Tpi1 antibodies. B, wild-type cells were grown in YPKG for 3d and transferred to YPD (left panels) or YP (right panels) for 0, 2, and 4 hours. Levels of Lia1p, Fbp1p, and Tpi1p were examined by Western blotting using anti-GFP, Fbp1p, and Tpi1p antibodies. C, wild-type cells were grown in YPKG for 3d and diluted to OD₆₀₀ = 0.2/ml in YPD (2% glucose), YPKG (0.5% glucose), or YP (0% glucose). Cell densities were measured at OD₆₀₀ for 0, 2, 4, 6, and 8 hours using a Beckman spectrophotometer.