FIGURE 2:

Protein stability of Tgl4p in cells defective in nonpolar lipid synthesis. Western blot analysis of Tgl4-Myc was performed with total cell extracts from wild-type (WT) (A), are1Δare2Δ (B), dga1∆lro1∆ (C), and QM (D) strains grown for times as indicated after addition of 100 μg/ml cycloheximide to cells grown to mid logarithmic phase. The primary antibody was directed against the Myc tag (Tgl4-Myc). The cytosolic marker GAPDH was used as loading control. Western blot analysis shown is one representative experiment of at least two independent experiments. (E) Relative amounts of Tgl4-Myc analyzed by Western blotting were calculated using ImageJ with the respective deviations as shown. The amount of Tgl4-Myc at time point 0 (addition of cycloheximide) was set at 1. WT, black bar; are1Δare2Δ, dark gray bar; dga1∆lro1∆, gray bar; QM, white bar.