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. 2005 Jan 20;24(3):580–588. doi: 10.1038/sj.emboj.7600540

Figure 2.

Figure 2

NBP35 is genetically linked to RIX19. (A) The NBP35 shuffle strain (nbp35∷KAN-MX4+pRS316-NBP35) was transformed with pRS315 plasmid containing nbp35-1 allele or the wild-type NBP35. After FOA selection, cells were grown on YPD plates for 5 days. (B) Rps2-GFP and Rpl25-GFP accumulation in the nbp35-1 ts mutant, which was transformed with indicated reporter plasmids. Exponentially growing cells were shifted from 30 to 37°C and examined after 4 h in the fluorescence microscope. (C) rix19-1 strain was crossed with the NBP35 shuffle strain. After tetrad analysis, a resulting rix19-1/nb35Δ haploid was transformed with NBP35 or nbp35-1. Two individual transformants were grown on the SDC plate containing 5-FOA for 5 days at 30°C. (D) The levels of ribosomal proteins Rpl10 and Rpl25 are not elevated in nbp35-1 strains. Overnight cultures of indicated stains were diluted to OD600 0.2 and grown at 23 or 37°C for 4 h. Whole cell lysates were separated by SDS–PAGE and transferred to nitrocellulose. Ponceau S staining (upper panel) revealed the protein loading, and the amount of Rpl10 and Rpl25 was determined by Western using specific antibodies (lower panel).