Figure 6.

Cell growth and expression of RanBP1 proteins in the rescued S. pombe sbp1-ΔC genomic mutant. (A) Growth of S. pombe haploid sbp1-ΔC genomic mutant cells transformed with nmt* plasmids encoding T7-tagged sbp1p, HA-tagged RBD (Sp), T7-tagged RanBP1 (m), or T7-tagged RBD (m) in liquid media in the presence (closed circles) or absence (open circles) of thiamine. Cell samples (20 μg of protein) collected at midlog phase when grown in the absence of thiamine or for an equal amount of time in its presence were analyzed by immunoblotting using anti-T7 or anti-HA epitope mAbs. (B) Growth of S. pombe haploid sbp1-ΔC and sbp1-Δ1 genomic mutants transformed with the plasmids encoding sbp1p, RBD (Sp), RanBP1 (m), or RBD (m) on Edinburgh minimal medium agar plates supplemented with adenine in the absence (MMA) or presence (MMAT) of thiamine and cultivated at 30° for 3 d.