Figure 8. XSox17α and XSox3 Bind to β-catenin and Interfere with Its Signaling (A) Secondary axis assay. mRNA encoding mutant stabilized pt-β-catenin (Yost et al. 1996) (0.1 ng), or pt-β-catenin (0.1 ng) + XSox17β, XSox17α, and XSox3 (0.5 ng) was injected into one ventral blastomere at the 4 cell stage. Tail bud stage embryos were scored for second axes. (B) OT transcription assay. 293T cells were cotransfected with the OT reporter construct (1 μg) and expression plasmids encoding S37A-β-catenin (0.3 μg), XSox17β-V5 (4 μg), XSox17α-V5 (4 μg), XSox3-V5 (4 μg), or S37A-β-catenin (0.3 μg) + each XSox17β-V5, XSox17α-v5, XSox3-V5 (2 μg and 4 μg; black triangles indicate increasing dose). (C) XSox17α and XSox3 bind β-catenin in vitro. 35S-labeled XSox17β, dXSox17β 1–150, XSox17α, and XSox3 proteins were incubated with 1 μg of GST-β-catenin fusion protein immobilized to agarose beads. Ten percent of the input proteins (i) and bound proteins (b). (D) XSox17α and XSox3 associate with β-catenin in vivo. 293T cells were cotransfected with S37A-β-catenin (2 μg) and expression plasmids (8 μg) encoding ΔN-hTCF4-V5 (lane 1), mLEF1-V5 (lane 2), hTCF4-V5 (lane 3), XSox17β-V5 (lane 4), XSox3-V5 (lane 5), or XSox17α-V5 (lane 6). The top panel shows an anti-V5 Western of the resulting extracts. Cell lysates were divided and subjected to immunoprecipitation with anti-β-catenin (+) or a negative control anti-mER antibody (−), followed by an anti-β-catenin Western to show levels of β-catenin (middle panel) and an anti-V5 epitope Western (bottom panel) to visualize proteins coprecipitating with β-catenin.
Image published in: Zorn AM et al. (1999)
Copyright © 1999. Image reproduced with permission of the Publisher, Elsevier B. V.
| Experiment + Assay | Source | Phenotypes and Disease | ||
|---|---|---|---|---|
| Xla Wt + ctnnb1tm1 + tailbud stage (morphology) | fig.8.a |
| ||
| Xla Wt + ctnnb1tm1 + hbp1 + tailbud stage (morphology) | fig.8.a |
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