Wnt3 adependent Cell Motility Involves Rho A Activation

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Wnt-3 a-dependent Cell Motility Involves Rho. A Activation and Is Specifically Regulated by Dishevelled-2

Wnt-3 a-dependent Cell Motility Involves Rho. A Activation and Is Specifically Regulated by Dishevelled-2 J. Biol. Chem. , Vol. 280, Issue 1, 777 -786, January 7, 2005

Cell line CHO Hamster Chinese ovary, Epithelial Migration assay Wound Healing Assay transwell assay

Cell line CHO Hamster Chinese ovary, Epithelial Migration assay Wound Healing Assay transwell assay

Wnt-3 a induced morphological changes in CHO cells Phalloidin staining

Wnt-3 a induced morphological changes in CHO cells Phalloidin staining

Wnt-3 a CM-stimulated migration of CHO cells. Wound Healing Assay (24 hr) transwell assay

Wnt-3 a CM-stimulated migration of CHO cells. Wound Healing Assay (24 hr) transwell assay (6 hr)

b -Catenin pathway was stimulated by Wnt-3 a CM but not required for migration

b -Catenin pathway was stimulated by Wnt-3 a CM but not required for migration in transwell assay.

Rho. A was activated by Wnt-3 a CM and contributed to Wnt-3 a-dependent cell

Rho. A was activated by Wnt-3 a CM and contributed to Wnt-3 a-dependent cell migration.

Wnt-3 a induced phosphorylation of Dvl-2 and Dvl-3

Wnt-3 a induced phosphorylation of Dvl-2 and Dvl-3

Intracellular distribution of Dvl-2 following treatment with Wnt-3 a or L CM. Wnt 3

Intracellular distribution of Dvl-2 following treatment with Wnt-3 a or L CM. Wnt 3 a CM L CM

Knock-down of endogenous Dvl-2 by RNAi inhibited Wnt-3 a-induced cell motility.

Knock-down of endogenous Dvl-2 by RNAi inhibited Wnt-3 a-induced cell motility.

GST-RBD pull-down assay the pull-down assay used to detect active small GTPases the N-terminal

GST-RBD pull-down assay the pull-down assay used to detect active small GTPases the N-terminal region of Raf 1 protein kinase contains the Ras–binding domain (RBD) the N-terminal regulatory region in p 21 -activated protein kinase 1 and 2 (Pak 1 and 2) contains the p 21 -binding domain (PBD) for Rac and Cdc 42, the N-terminal region of Rhotekin contains the Rho-binding domain (RBD). The GTPase-binding domains from these downstream effectors are expressed as recombinant glutathione S-transferase (GST) fusion proteins immobilized on glutath and can be used to affinity precipitate (pull-down) the active GTPase from cell lysates. Pulled-down active GTPases are eluted from the resin and detected by immunoblotting with a specifi