Skip to main content
. Author manuscript; available in PMC: 2010 Nov 13.
Published in final edited form as: Mol Cell. 2009 Nov 13;36(3):525–535. doi: 10.1016/j.molcel.2009.10.025

Fig. 5.

Fig. 5

Strategy for the simultaneous, localized manipulation of PI(4,5)P2- and Nck-dependent signaling at the plasma membrane. A) Diagrams representing the domain structure of Nck, the PI 5-phosphatase synaptojanin-1, and the fusion of CD16/7 with all three Nck SH3 domains (CD16/7-Nck), or the PI 5-phosphatase domain (CD16/7-SJ). The fusion proteins were tagged with the HA epitope. B) Diagram illustrating the antibody-mediated clustering utilized to simultaneously increase the local concentration of membrane-targeted Nck SH3 domains and decrease the local concentration of PI(4,5)P2. C) Expression of CD16/7-SJ (wild type) and CD16/7-D858A (catalytically inactive) in NIH3T3 cells demonstrated by western immunoblotting with anti-HA. D) Confocal images of NIH3T3 cells expressing CD16/7-Nck (CD16/7-Nck, left panel), or a fusion of CD16/7 with the wild type IP 5-phosphatase domain of synaptojanin-1 (CD16/7-SJ) following treatment with clustering antibodies (A647). Scale bar represents 5 μm.