XB-IMG-83468
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Figure S2. RAB8A Knockdown Has Weak or No Phenotype in Wnt Assays, Epistasis Analysis of RAB8B, and Depletion of RAB8B Reduces Wnt Activity in Wnt-Inducible HeLa Cells, Related to Figures 1 and 2
(A) RAB8A knockdown showed a weak phenotype in Wnt reporter assay in HCT116 and HEK 293T cells.
(B and C) RAB8A silencing did not change Wnt target gene AXIN2 levels and cellular viability in HCT116.
(D) RAB8B siRNAs reduced Wnt reporter activity when pathway was stimulated with Wnt3a coexpression but not with Xenopus tropicalis β-catenin coexpression in HEK 293T.
(E) LRP6 RNAi reduced target gene AXIN2 levels one-third with a mild knockdown efficiency (HCT116).
(F) RAB8B siRNA#1 reduced AXIN2 mRNA levels after 96 hr of incubation (HCT116).
(G) Wnt target genes were induced using Wnt conditioned medium in HeLa cells. RAB8B RNAi reduced AXIN2 levels activated with 3 hr of Wnt3a exposure.
(H) Active β-catenin accumulation was impaired in RAB8B depleted HeLa cells that were activated with Wnt3a exposure for 3 hr.
(I) Quantification of nuclear β-catenin in Figure S2H was performed with ImageJ.
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. Larger Image Printer Friendly View |