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. 2015 Aug 3;210(3):471–483. doi: 10.1083/jcb.201411033

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© 2015 Lin et al.

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Figure 7. — Kinase activity of Lrrk proteins in Lva phosphorylation and Dhc interaction. (A and B) Western blots show pulled-down Dhc (A) or Lrrk (B) by GST-Lva5 or GST-Lva1 from fly head extracts of elav-GAL4 control and Flag-Lrrk or Lrrk-3KD overexpression. GST as pull-down controls and α-Tub as input controls. (C) Western blot shows pSer/Thr of Lva in Lva immunoprecipitates for +/del6, e03680/del6, and Flag-Lrrk or Lrrk-3KD overexpression by elav-GAL4. (D–F) Western blots show coprecipitated pSer/Thr and GST (D), Lrrk2 and DHC (E) or Lrrk2, DHC, pSer/Thr, and GST (F) in GST pull-down complexes from incubations of Lrrk2 with GST-Lva5 (D) or with GST-Lva5 and dynein (E and F) with or without 10 mM ATP followed by GST pull-down. Input control was analyzed by antibodies for Lrrk2, DHC, and GST (F). Experiments were all performed three times except once for C because of a limited amount of Lva antibodies.