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. 1999 Aug 3;96(16):9112–9117. doi: 10.1073/pnas.96.16.9112

Figure 3.

Figure 3

Nuclear export by Crm1-K1 is resistant to LMB and NEM. S. pombe strains JY266 (crm1+) and KY21 (crm1-K1) were transformed with pR1GrevNESF1 for GST-Rev NES-GFP (GST-NES-GFP) and pR1GsvNLSFrevNES1 for GST-SV40 T NLS-GFP-Rev NES (GST-NLS-GFP-NES), as described (9). These genes are under the control of the wild-type nmt1 promoter, which can strongly direct transcription when thiamin is absent in the medium. Cells were cultured in the absence of thiamin for 12 h at 30°C for induction of the proteins and then cultivated further for 6 h in the same medium with or without LMB (50 ng/ml) or NEM (100 μM) at 30°C. Cells were stained with 4′,6-diamidino-2-phenylindole for DNA and observed under a Zeiss Axiophot-2 microscope.