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. 2009 Jun 5;106(25):10183–10188. doi: 10.1073/pnas.0812105106

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Fig. 1. — GRK2 interacts with PTCH1. (A) Interaction between PTCH1 and GRK2 was detected in HEK293 cells transfected with human myc-tagged Patched 1 (PTCH1-myc) and/or bovine Flag-tagged GRK2 (GRK2-Flag) cDNA. Lysates were immunoprecipitated with anti-Flag antibody (B). The immunocomplex (Upper) and the input cell lysate (Lower) were detected by Western blotting using the indicated antibodies. (B) Lysates of nontransfected HEK293 cells were immunoprecipitated with anti-PTCH1 antibody (Santa Cruz). (C) Direct interaction of GRK2-Flag with GST-tagged segments of PTCH1, GST-PTCH1-N (1–502 amino acids), GST-PTCH1-M (503–1121 amino acids), and GST-PTCH1-C (1180–1447 amino acids) was examined in GST-pull down assays. (D and E) Interactions between PTCH1 and GRK2 mutants Δ245–312, Δ312–379, Δ379–453, Δ453–512 (D) and 262–379 (BP-Flag) (E).