FIGURE 8:

Protein stability of Tgl3p, Tgl4p, and Tgl5pin the absence of counterpart lipases. Western blot analysis of (A) Tgl3-Myc in tgl4∆tgl5∆, (C) Tgl4-Myc in tgl3∆tgl5∆, and (E) Tgl5-Myc in tgl3∆tgl4∆ was performed with total extracts from cells 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 (Tgl3-Myc, Tgl4-Myc, and Tgl5-Myc). The cytosolic marker GAPDH was used as loading control. Western blot analyses shown are one representative experiment of at least two independent experiments. Relative amounts of Tgl3-Myc (B), Tgl4-Myc (D), and Tgl5-Myc (F) obtained by three Western blots were calculated using ImageJ with the respective deviations as shown. Amounts of the respective protein at time point 0 (addition of cycloheximide) were set at 1. WT, wild type.