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. Author manuscript; available in PMC: 2008 Aug 24.
Published in final edited form as: J Biol Chem. 2007 Jun 12;282(34):24743–24751. doi: 10.1074/jbc.M700663200

Figure 1. Assembly properties of subunit a isoforms localized to their normal cellular environment.

Figure 1

Yeast strain MM112 (vph1Δ stv1Δ) containing plasmids pSKN12 (pRS316-VPH1::HA) or pSKN11(pRS316-STV1::HA) were grown to log phase, harvested, and converted to spheroplasts with Zymolyase. Whole cell lysates were prepared by treatment with C12E9 (with or without 1 μM concanamycin) and V0 complexes were immunoprecipitated using the antibody 3F10 (Roche) against the HA epitope. Following SDS-PAGE, Western blotting was performed using the antibody 8B1-F3 (Molecular Probes) against subunit A (Vma1p) as well as 3F10 against HA. The amount of subunit A co-precipitated with the antibody against HA is a reflection of the degree of assembly of V1 and V0.