Localization of Mig2-GFP in kap95tsand kap60tsyeast cells. (a) The FMY527 (kap95ts Mig2-GFP) and (b) FMY528 (kap60ts Mig2-GFP) strains were transformed with plasmid pRS316/Su9-RFP. Transformed cells were grown in high-glucose synthetic medium (H-Glc) until an A600nm of 1.0 was reached and then transferred to low glucose synthetic medium (L-Glc) for 5 min. (c) The JCY1410 (kap95ts) strain transformed with plasmid YEp352/Hxk2nes2(Ala)-GFP was used as positive control. Transformed cells were grown in high-glucose synthetic medium (H-Glc) until an A600nm of 1.0 was reached. The cells were grown at 25°C (permissive temperature) and then shifted to 37°C (not permissive temperature) for 1 h. Cells were stained with DAPI and imaged for GFP, RFP and DAPI fluorescence. Scale bar is 10 μm. The nuclear or mitochondrial localization of Mig2-GFP and the nuclear or cytoplasmic localization of Hxk2nes2(Ala) proteins was determined in at least 100 cells per growth condition. Error bars represent standard deviations for three independent experiments. N denotes a nuclear fluorescence signal and M mitochondrial fluorescence signal.