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. Author manuscript; available in PMC: 2013 Jul 30.
Published in final edited form as: FEBS Lett. 2012 Jun 20;586(16):2294–2299. doi: 10.1016/j.febslet.2012.06.016

Figure 5. Interaction between paxillin and β-catenin is Rac dependent.

Figure 5

A: HEK293T were co-transfected with full length GST-tagged paxillin or its domains, HA-tagged dominant negative (Rac1-N17) or constitutively active (Rac1-V12) Rac mutants, and full length His-tagged β-catenin. After 48 hrs of transfection, the cells were stimulated with vehicle or EGF (500 ng/ml, 15 min) and lysed. Pulldown assays on GSH beads were performed as described in Materials and Methods. The content of β-catenin bound to GST-paxillin or its domains was determined by western blot analysis with corresponding antibody (upper panel). Expression levels of GST-tagged paxillin mutants were detected in whole cell lysates using GST antibody (lower panel). B: Quantification of pulldown experiments. Amount of β-catenin bound to GSH beads with paxillin or its domains was determined by quantitative densitometry of western blot membranes and normalized to the amount of immobilized paxillin and its domains on the beads. Results of densitometry are expressed as β-catenin/GST ratio and shown as mean ± SD, * p<0.05 vs. non-stimulated control; n=4.