Glucose-dependent degradation of Mth1 takes place in the nucleus. Synthetic oligonucleotides encoding the nuclear export sequence (NES) and its mutant peptide (NES (m)) were inserted between the GFP and MTH1 genes in the GFP-MTH1 construct. Yeast cells (mth1Δ, YM6266) expressing GFP-Mth1 and its derivatives were grown as described in Figure 2. Subcellular localization and levels of GFP-Mth1 were analyzed by fluorescence microscopy using CLSM (A) and Western blotting using anti-GFP antibody (B), respectively.
Pasula et al. BMC Cell Biology 2010 11:17 doi:10.1186/1471-2121-11-17