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. 2010 Sep 29;5(9):e13064. doi: 10.1371/journal.pone.0013064

Figure 3. Interaction of DAAM1 with Rho GTPases.

Figure 3

(A) The design of putative Rho binding defective mutants was based on sequences in mDia1: RhoA complex [6]. An in-vitro pulldown assay was performed in which equal amounts of DAAM1-N wildtype (WT) or the GBD mutants were added to Sepharose beads loaded with GST or GST-RhoAV14. The proteins were transferred to PVDF and stained as shown. The membrane was subsequently probed as indicated (B) GFP-DAAM1-N WT or the GBD mutants were expressed in COS-7 cells and immuno-localized with anti-GFP and F-actin (TRITC-phalloidin); all three constructs show typical DAAM1 filamentous staining. Bars  = 10 µm.