Fig. 3. Cln2 localizes to the cytoplasm and to the nucleus, and the forced localization cassettes can shift Cln2 to specific compartments. (a) GFP-CLN2-18xMYC (plasmid pMGH101) was introduced to strain N-104(#31) (cln1 cln2 GAL-CLN3 cdc34-2), which also contains a centromeric plasmid containing CLN3 under its own promoter (pHW415). Cells were grown under selective conditions (–met 2% glucose) at 25°C to early log phase, and then shifted to 37°C for 1-3 hours. Live cells were stained with Hoechst 333642, mounted under coverslips in mounting media, and images were captured digitally. The cell on the right rotated slightly between imaging. (b) Plasmids (pMGH10X series) containing GFP-CLN2 fused to the forced localization cassettes at the Cln2 C-terminus were introduced to strain N-104(#31) and analyzed as described in (a).