Figure 5.

Analysis of the stability of the Mdm34, Mdm12 and mutant Mdm34[3PA]proteins. WT and rsp5Δ pspt23ΔCT strains bearing chromosomal genes encoding 9MYC-tagged MDM34, mdm34[3PA], or MDM12 were grown to mid-exponential growth phase in YPD at 24°C. The cells were then shifted to 37°C, and protein synthesis was blocked by adding cycloheximide (CHX, 200 µg/mL). Protein extracts were prepared at the times indicated and analyzed by western immunoblotting with an anti-MYC antibody. Pgk1 was used as a loading control.