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. 2009 Jan 1;20(1):153–163. doi: 10.1091/mbc.E08-02-0157

Figure 6.

Figure 6.

TAP tag pull-downs of the exocyst by using either Exo70-TAP or Exo70ΔdC-TAP show reduced levels of Sec6p in the assembled complex. The negative control lane (NEG) is a pull-down from a strain containing 3xHA-tagged Sec5p but no Exo70-TAP or Exo70ΔdC-TAP. (A) Silver-stained gel of the isolated exocyst complex from either wt (EXO70) cells or exo70ΔdC (ΔdC) cells. Exo70-TAP or Exo70ΔdC-TAP was pulled down, and the resulting complexes treated with TEV protease to release the exocyst complex from the IgG beads. The released exocyst complexes were then isolated on calmodulin beads and separated by SDS-polyacrylamide gel electrophoresis for silver staining. Exo70-CBP and exo70 ΔdC-CBP indicate Exo70p and Exo70 ΔdCp, respectively, with the calmodulin binding peptide after TEV cleavage. (B) Western blots detecting different exocyst components in exocyst complexes isolated from Exo70-TAP and Exo70ΔdC-TAP pull-downs. The input lanes represent 1% of total protein, and the blot was probed with anti-HA antibodies to 3xHA-tagged Sec5p.