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. 2005 Jun;25(11):4476–4487. doi: 10.1128/MCB.25.11.4476-4487.2005

FIG. 5.

FIG. 5.

Rmi1 binds Top3 and Sgs1 in recombinant form. Cell lysates were prepared from bacterial strains expressing the indicated epitope-tagged proteins in the left column (Extract). Extracts were immunoprecipitated with the indicated antibodies, washed three times in RIPA buffer, and immunoblotted with either anti-HA (left) or anti-V5 (right). (A and B) Rmi1-HA and Top3-V5 were coexpressed from plasmid pCS7141. (C and D) Rmi1-HA and Sgs1-V5 were coexpressed from plasmid pNJ1572. (E and F) Sgs1-V5, untagged Top3, and Rmi1-HA were coexpressed from plasmid pNJ1571. (G) Rmi1-HA was expressed alone from plasmid pNJ7135. EXT is a control representing 1/20 of the amount of extract subjected to IP. ΔSgs1-V5 indicates the position of a 150-kDa breakdown product of full-length Sgs1-V5. Molecular mass markers are shown to the left of each blot in kilodaltons.