Figure S3. Synergistic Phenotype of RAB8B Is Conserved, and Further Confirmation of Dvl1-RAB8B Colocalization, CK1γ Rescues RAB8B Knockdown, and Control Co-IP with FLRT3, Related to Figure 3 (A) Xenopus laevis rab8b-c.a. synergized with LRP6, and further induced Wnt reporter activity (HEK 293T). (B) RAB8B-w.t. expression failed to synergize with LRP6 in the Wnt reporter assay (HEK 293T). (C) In Wnt active HCT116 cells, both RAB8B-w.t. and c.a synergized with LRP6 and further activated Wnt pathway, while d.n. form did not. (D) RAB8B-w.t. further induced pathway activity only with Dvl1 coexpression (HEK 293T). (E) Dvl1 coexpression recruited RAB8B in large Dvl1 puncta (HeLa). (F) Dvl1 antibody gives a nonspecific nuclear staining in the nucleolar region as it is persistent in Dvl1 siRNA transfected cells. Cyoplasmic staining of Dvl1 is specific since it is induced by Dvl1 coexpression and can also be depleted by Dvl1 siRNA. (G) Dvl1 knockdowns are efficient (HeLa). (H) RAB8B knockdown reduced Wnt reporter activity induced by LRP6 coexpression, and this phenotype was rescued with coexpression of CK1γ (HEK 293T). (I) RAB8B coimmunoprecipitated with LRP6 but not with a non-Wnt related transmembrane protein FLRT3 (HEK 293T). Error bars represent �SD of three replicates.
Image published in: Demir K et al. (2013)
Copyright © 2013. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.
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