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. 2007 Jul 16;178(2):297–308. doi: 10.1083/jcb.200701078

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Copyright © 2007, The Rockefeller University Press

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Figure 4. — PIPKIγ is required for EGF-induced talin assembly to adhesions. (A) The expression of PIPKIγ668 in control shRNA– or PIPKIγ668-specific shRNA–transfected HeLa cells was detected. (B) Control shRNA or PIPKIγ668-specific shRNA–transfected HeLa cells were stimulated with 10⁻¹⁰ M EGF for 15 min. Cells were fixed and stained with anti-talin antibody. (C) EGF-induced talin assembly to adhesions was quantified. (D) Control shRNA or PIPKIγ668-specific shRNA were cotransfected with GFP-talin into HeLa cells. A micropipette filled with EGF was put near the cell to stimulate GFP-talin recruitment. GFP-talin assembly kinetics was quantified. Rate constants for assembly of individual adhesions were calculated as described in Materials and methods. Quantifications are expressed as means ± SEM (*, P < 0.01 compared with control siRNA–transfected HeLa cells). (E) GFP-talin assembly to adhesions at different time point of micropipette stimulation was shown. See Videos 1 and 2 (available at http://www.jcb.org/cgi/content/full/jcb.200701078/DC1). Bars, 10 μm.