Figure 9.

Cell growth, protein expression, and nuclear import in the S. pombe sbp1-ΔC genomic mutant rescued with RBDs 2 and 4 of RanBP2. (A) Growth in liquid media in the presence (repressed, closed circles) or absence (derepressed, open circles) of thiamine and detection by immunoblotting of HA-tagged proteins present in 20 μg of total cell extracts. sbp1-ΔC was rescued with HA-tagged RBD2 or RBD4 expressed from full-strength nmt promoter plasmids. (B) Immunodetection of HA-tagged RBD2 and RBD4 of RanBP2, and DAPI staining of cellular DNA, in cells grown in the presence or absence of thiamine. (C) GST-GFP-NLS localization in sbp1-ΔC cells rescued by RBD2 or RBD4, grown in the presence or absence of thiamine, and analyzed as described in the legend to Figure 8.