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. 2010 Jul 20;29(17):2899–2914. doi: 10.1038/emboj.2010.159

Figure 8.

Figure 8

Differential binding of cytosolic and PM-associated Myo5 to Cmd1 and Vrp1. (A, C) Immunoblots of 10 μg of total (T), PM or cytosolic (C) fractions prepared from myo5Δ cells (SCMIG275) (A) or myo5Δ vrp1Δ cells (SCMIG304) expressing Vrp1-HA (C), expressing the indicated PA-Myo5 constructs. PAP, a peroxidase-conjugated anti-HA antibody and antibodies against Cmd1 or the PM and cytosolic markers Gas1 and Hxk1, combined with the appropriate peroxidase-conjugated secondary antibody, were used for detection of the PA-Myo5 constructs, Vrp1-HA, Cmd1, Hxk1 or Gas1, respectively. (B, D) Immunoblots of IgG pull downs from yeast PM and cytosolic (C) fractions from myo5Δ cells (SCMIG275) (B) or myo5Δ vrp1Δ cells (SCMIG304) expressing Vrp1-HA (D), expressing the indicated PA-Myo5 constructs. Membranes were processed as described in (A, C).