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. 2013 Feb 25;8(2):e57001. doi: 10.1371/journal.pone.0057001

Figure 7. TbMCM-BP co-purifies with MCM4-MCM8.

Figure 7

(A) TbMCM-BP interacts with MCM5 but not with MCM2 by co-immunoprecipitation. (B) Allele modification in cells exclusively expressing TbMCM-BP tagged with PTP. (C) Immunoblot monitoring of TbMCM-BP-PTP tandem affinity purification in extract (Input), IgG affinity chromatography flowthrough (FT-IgG), TEV protease eluate (Elu TEV), flowthrough of the anti-ProtC affinity chromatography (FT-ProtC), and final eluate (Elu). Note that TEV protease-mediated removal of the ProtA domains led to a size reduction of tagged TbMCM-BP (‘BP-PTP’ to ‘BP-P’). Relative amounts of aliquots are specified by x values. (D) Tandem affinity purification of TbMCM-BP. Final eluate and smaller aliquots of the TEV protease eluate and extract were separated on an SDS–gradient polyacrylamide gel and stained with Coomassie blue or Sypro Ruby. (E) TbMCM-BP is localized in the nucleus throughout the cell cycle in PF.