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Fig. 3. CrkII forms a Reelin-promoted complex with Dab1 in embryonic forebrain neurons. Total cell lysates (TCL) were collected from cultured embryonic forebrain neurons which were unstimulated (0; lanes 1,4), stimulated with control conditioned media (CCM; lanes 2,5), or stimulated with Reelin conditioned media (RCM; lanes 3,6) for 15 minutes. Western blotting these samples with anti-phosphotyrosine antibody (pY; 4G10) shows the induction of the tyrosine phosphorylation of an 80 kDa protein, which corresponds to Dab1 (lanes 1-3, upper panel), whereas the Dab1 protein levels are relatively constant between samples (lanes 1-3, lower panel). Equal levels of CrkII are detected in the anti-CrkII immunoprecipitates from these cell lysates (lanes 4-6, upper panel); however, increased co-immunoprecipitation of Dab1 was detected from lysates of Reelin-treated samples (compare lanes 4-6, lower panel). Approximately 100% of CrkII was immunoprecipitated under the assay conditions, and 5% of total Dab1 was detected in the co-immunoprecipitation (not shown).





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