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. 2009 Feb 5;1(2):219–233. doi: 10.18632/aging.100020

Figure 5. WRN mediated restoration of slow growth phenotype in sgs1 top3 background is independent of its expression level.

Figure 5.

WRN expression in the transformed sgs1 top3 cells was induced with the indicated gal concentrations and cells were harvested 6 h after induction. As a control, sgs1 top3/YEp112SpGAL was included. Equal amounts of total cell lysate were loaded on 8-16% polyacrylamide SDS gels followed by Western blot detection using anti-WRN antibody as shown in Panel A. sgs1 top3 strain transformed with YEp112SpGAL, YEp112SpGAL-WRN, exonuclease-dead (YEp195SpGAL-WRN E84A), ATPase/helicase-dead (YEp195SpGAL-WRN K577M), RQC mutant (YEp195SpGAL-WRN K1016A), polymorphic mutant(YEp195SpGAL-WRN R834C) and YEp112SpGAL-SGS1 was streaked on SC-Trp plate containing gal at varying concentrations asindicated. Plates were incubated at 30°C for 2 days and then photographed. Panels C-F show the effect of WRN expression onthe growth rate of sgs1 top3 transformed strains at different gal concentrations. Composition of the plates was as in Panel B.