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. 1998 Apr 28;95(9):5051–5056. doi: 10.1073/pnas.95.9.5051

Figure 2.

Figure 2

PYK2 points mutations identify several events regulated by the m1 receptor. (A) Stable cell lines expressing the m1 receptor together with HSV-tagged PYK2, kinase inactive PYK2-K457A, PYK2-Y402F, or PYK2-Y881F were treated for 2 min with vehicle (−) or 50 μM carbachol (+). Lysates from these cell lines were immunoprecipitated with anti-HSV antibody and determined by Western blot analysis with either anti-phosphotyrosine antibody (Upper) or anti-HSV antibody (Lower). (B) Anti-HSV immunoprecipitated lysates of the indicated cell lines prepared before (−) and after (+) carbachol treatment (2 min, 50 μM) were immunoblotted with antibodies against c-Src (Upper Left) or Grb2 (Upper Right). In each case the level of PYK2 present in immunoprecipitates was assessed by Western blot analysis with anti-HSV antibody (Lower). (C) Cells expressing PYK2-HSV were lysed before (−) and after (+) a 2 min 50 μM carbachol stimulation. Anti-HSV immunoprecipitates from the lysates were analyzed by Western blot using anti-paxillin (Upper Left) or anti-HSV (Lower Left) antibodies. Anti-paxillin immunoprecipitates were analyzed by Western blot using anti-phosphotyrosine antibodies (Upper Right) or anti-paxillin (Lower Right).