Figure 4.—

Perinuclear localization is required for repression by Mig1. (A) Confocal laser scanning microscopy of Mig1-GFP in wild-type or Δhxk2 cells grown in either the presence or the absence of glucose, as indicated; coexpressed Rap1-CFP is shown as a nuclear marker. GFP and CFP signals were captured by using a Zeiss LSM 510 META confocal laser scanning microscope with a 63× Plan-Apochromat 1.4 NA oil DIC objective lens. Signals were separated with a 490-nm dichroic mirror with long pass filters adjusted to 505 and 475 nm, respectively. Pinholes were adjusted to obtain <1.8-μm optical slices. Images were acquired with the Zeiss LSM 510 software version 3.2. (B) Graphs show the fraction of total Mig1-GFP fluorescence present in cytoplasmic (Cyto), nucleoplasmic (Nuc-P, total nuclear fluorescence minus perinuclear fluorescence), and perinuclear (Perinuc) fractions, as determined by QFPD analysis. Open bars, glucose; shaded bars, no glucose. Error bars represent the standard error of the mean.