Xenbase Image ID: 83610

Figure S3. PACSIN2 does not bind α5β1. (A) Immunoprecipitations of protein extract of 10 gastrulas was performed using mAb against PACSIN2 (3D8), and α5β1 (P8D4). The precipitates were analyzed by western blot using biotinylated mAb against PACSIN2 (3D8biot, left panel). The blots were stripped in 200 mM glycin pH 2.8 and re-blotted with a rabbit polyclonal antibody against α5 integrin (881, right panel) (Joos et al. 1995). The P8D4 antibody successfully precipitated the α5β1 integrin but failed to co-immunoprecipate PACSIN2. Note that the PACSIN2 signal was incompletely stripped and therefore appear in the integrin β1 blot. (B) Pull Down experiment was performed using protein extract of 10 gastrula and 1μg of GST alone or GST fusion proteins of integrin α5 or β1 tail. The proteins bound were analyzed by western blot using the PACSIN2 mAb (3D8). The protein extract of 1 embryo equivalent was loaded as a positive control (extract). A ponceau staining of the membrane is shown on the right. The fusion protein corresponding to the Image published in: Cousin H et al. (2008) Copyright © 2008. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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