Figure 3.—
Nuclear export of Ltv1-GFP is Crm1 dependent. (A) pLTV1-GFP (Ld47), pYAR1-GFP (Ld46), and NLSSV40-NESPKI-GFP (pPS1372) plasmids were transformed in the LMB-sensitive crm1(T539C) (MNY8) strain. Strains expressing these GFP-tagged proteins were grown overnight at 30° in appropriate selective media to early to midlog phase, harvested, and photographed as described in materials and methods. LMB was added to an aliquot of each culture to a final concentration of 100 ng/ml and images of living cells captured beginning 5 and 60 min after addition of LMB. Time, in the left-most column, is the time since addition of LMB to cells. (B) Living cells expressing Ltv1-GFP (Ld47) or NLSSV40-NESPKI-GFP (pPS1372) protein were stained with DAPI and then treated with LMB. Cells were photographed 5 min after the addition of LMB.