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. Author manuscript; available in PMC: 2011 Feb 1.
Published in final edited form as: Traffic. 2009 Nov 19;11(2):185–197. doi: 10.1111/j.1600-0854.2009.01010.x

Figure 4.

Figure 4

Chz1 contains an NLS and associates directly with Srp1/Kap95 in cytosol. (A) Fragments of Chz1, bearing the indicated mutations, fused to GFP2 were expressed in wild type yeast as indicated and visualized by fluorescence microscopy. Coincident Hoechst staining is shown. (B) Chz1-TAP was immunoprecipitated from cytosol prepared from strains expressing also Kap95-Myc or Kap114-Myc. Bound protein was eluted with MgCl2, precipitated, and analyzed by SDS-PAGE and Western blotting (WB) using anti-Myc antibodies. Chz1-TAP is recognized by IgG. `Un' lane contains 0.02% of depleted post-bead starting cytosol. (C) Recombinant H6-PrA-Chz1 (250 nM) was immobilized on IgG sepharose and incubated with H6-Srp1 (500 nM) and Kap95 (500 nM). zz protein (~250 nM) was used as a negative control. Bound protein was eluted and analyzed by SDS-PAGE and Coomassie blue staining (CBB).