Rack1 interacts with Vangl2. (A) A diagram depicting the use of Vangl2c as the bait to screen a cochlear cDNA library to identify cytoplasmic Vangl2-interacting proteins. Yeast cells were serially diluted and replicated on rich media YAPD (yeast media containing yeast extract, peptone, dextrose plus adenine) plates and minimal select plates. (B) Vangl2EGFP and Rack1 can be coimmunoprecipitated with embryonic day 16.5 mouse embryo brain and cochlear extracts prepared from transgenic mice expressing Vangl2-GFP.(C) Rack1WD5–7 was efficiently pulled down by Vangl2, and there are two bands for HA-Rack1. The rabbit polyclonal antibody against Rack1 also recognizes two bands for the endogenous Rack1 (C), whereas the monoclonal antibody recognizes one band (B).
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