XB-IMG-176107
Xenbase Image ID: 176107
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Fig. 3. PTK7 cPTK7 622–1070 and 726–1070 proteins support LRP6 activation of Wnt direct-target genes
A. Embryos were injected at the one-cell stage with PTK7-MO (10 ng, lanes 5–7). In parallel, these embryos co-injected animally with mRNA encoding the ligand-independent form of the LRP6 protein, LRP6ΔN (175 pg) that activates the Wnt pathway (lane 4), and mRNA encoding the full-length PTK7 protein (200 pg, lane 7). Animal caps (ACs) removed at the blastula stage were grown until gastrula st.10.5, when total RNA was isolated from five control embryos (lane 2) and 18 ACs from each group (lanes 3–7). Wnt direct-target gene expression was examined by sq (semiquantiative) RT-PCR: xnr3, siamois and hoxd1. Ef1α serves as positive control for RNA levels.
B. Embryos were injected at the one-cell stage with PTK7-MO (10 ng, lanes 5–8). In parallel, these embryos co-injected animally with mRNA encoding the ligand-independent form of the LRP6 protein, LRP6ΔN (175 pg) that activates the Wnt pathway (lane 4), and mRNA encoding the short PTK7 proteins cPTK7 622–1070 and cPTK7 726–1070 (200 pg, lanes 7–8). ACs removed at the blastula stage were grown until gastrula st.10.5, when total RNA was isolated from five control embryos (lane 2) and 18 ACs from each group (lanes 3–9). Wnt direct-target gene expression was examined by sqRT-PCR: xnr3, siamois and hoxd1. Ef1α serves as positive control for RNA levels
C. Embryos were injected at the one-cell stage with PTK7-MO (12 ng, lanes 4–6). In parallel, these embryos co-injected animally with mRNA encoding LRP6 protein (600 pg, lane 3), and mRNA encoding the short PTK7 proteins cPTK7 622–1070 and cPTK7 726–1070 (200 pg, lanes 5–6). Embryos were grown until gastrula st.10.5, when total RNA was isolated from five embryos in each group. Wnt direct-target gene expression was examined by sqRT-PCR: hoxd1 and gbx2. Siamois expression is unchanged as described in text. Ef1α serves as positive control for RNA levels. Image published in: Lichtig H et al. (2019) Copyright © 2019. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View |