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. 2001 May 1;15(9):1104–1114. doi: 10.1101/gad.879301

Figure 1.

Figure 1

Subcellular localization of Sip1, Sip2, and Gal83. Strains MCY2649, MCY2700 (sip2Δ), and MCY4458 (gal83Δ) were transformed with centromeric plasmids expressing Sip1–GFP, Sip2–GFP, or Gal83–GFP, respectively; plasmids were pOV90, pRT9, and pRT12. (A) Cells were grown in synthetic medium with 2% glucose (Glu) or 2% glycerol plus 3% ethanol (G + E). Protein extracts were prepared, separated by SDS-PAGE, and immunoblotted with anti-GFP antibody. (B) The subcellular localization of each GFP fusion protein was examined by fluorescence microscopy in cells grown on synthetic medium with 2% glucose (Glu), 2% glycerol plus 3% ethanol (Gly + Eth), or 5% glycerol (Gly). The arrows indicate that cells were shifted from the growth medium to a different carbon source and incubated for 20 min before observation. Cells grown in glucose were shifted to 2% or 5% glycerol, and cells grown in glycerol were shifted to 5% glycerol plus 2% glucose. Nuclear export of Gal83 also was observed when cells were shifted from 5% glycerol to 2% glucose (data not shown). A low level of autofluorescence was observed with untransformed strains (not shown). DNA was stained with DAPI. (GFP) green fluorescent protein.