XB-IMG-49522
Xenbase Image ID: 49522
|
Figure 4. b-Catenin Is Required for GSK3 Localization in Multivesicular Endosomes
(A and B) b-Catenin siRNA inhibited GSK3b relocalization in CA-LRP6 signalosomes (in 86% 7% of transfected HeLa cells, n = 300).
(C) HRSMOblocks the induction of TCF reporter expression (brackets) by b-Catenin mRNA (four injections of 80 pg) in Xenopus animal cap explants, and this was
partially rescued by human HRS mRNA.
(D0 ) Endogenous phospho-b-Catenin colocalizes with CA-LRP6-GFP signalosomes in HeLa cells (85% 3% of transfected cells, n = 56).
(G0 ) Overexpression of a stabilized mutant of b-Catenin-GFP caused its accumulation both in the nucleus and in cytoplasmic particles that sequester GSK3-RFP
from the cytoplasm (75% 11%, n = 60).
(J0 ) Wild-type b-Catenin-GFP becomes localized inside giant endosomes induced by Rab5-QL-DsRed. Seventy-nine percent 10% of giant MVBs contained
b-Catenin-GFP, n = 80.
(M) Axis induction by b-Catenin mRNA (80 pg) in Xenopus was blocked by coinjection of HRS-MO and partially rescued by 10 pg human HRS mRNA.
(P) Vps4-EQ, but not Vps4-WT, mRNA inhibited secondary axis formation by b-Catenin mRNA.
(S) Nuclear function of b-Catenin in Xenopus embryos. Wild-type, but not b-Catenin depleted embryos, contains neural tissue marked by Sox2 (n = 44 and
n = 25). Microinjection of DN-GSK3 mRNA (150 pg, four times at four-cell) dorsalized in a b-Catenin dependent manner (n = 38 and n = 28). DN-Tcf3 mRNA
(200 pg, four times) blocked dorsalization by DN-GSK3 (n = 36), whereas the fusion construct b-Catenin-DN-xTcf3 (30 pg) rescued the ventralizing effects of
b-Catenin MO (n = 26). Arrowheads indicate position of the blastopore.
(Ya) Epistatic experiment showing that b-Catenin-DN-xTcf3 fusion protein does not require MVB formation to induce secondary axes in Xenopus embryos.
(Baa) Epistatic experiment showing that the downstream target of Wnt signaling Siamois is not affected by MVB inhibition.
Data are represented as mean SEM. Also see Figure S5. Image published in: Taelman VF et al. (2010) Copyright © 2010. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View |