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Fig. 7. Constitutively activated LRP6 proteins interact with endogenous LRP6 protein. (A) Embryos were injected at the one-cell stage with RNA encoding LRP6δN (170 pg) or myr-LRP6-ICD (500 pg) proteins (constitutively activated constructs) and the PTK7-MO (10 ng) or LRP6-MO (10 ng). AC explants were removed from control and injected embryos at blastula stage and grown to gastrula st.10.5. Total RNA was isolated from six control embryos and 27 ACs from each group. Wnt direct-target gene expression was examined by sqRT-PCR. (B) Embryos were injected at the one-cell stage with myr-LRP6-ICD mRNA (1 ng) and the PTK7-MO (10 ng), LRP6-MO (20 ng) or both. ACs removed at the blastula stage were grown to gastrula st.10.5. Total protein was isolated from 27 ACs per group for western analysis using the anti-LRP6 T1479 antibody. (C) Embryos were injected at the one-cell stage with RNA encoding LRP6δN (100 pg) or myr-LRP6-ICD (400 pg) proteins and the PTK7-MO (13 ng). ACs were cultured as described, and lysis was performed on 36 pooled explants per AC group for sqRT-PCR. Wnt direct-target gene expression was examined by sqRT-PCR. (D) Duplicate western analysis of the experiment described in C. For western analysis, the T1479 antibody recognizes LRP6δN and myr-LRP6-ICD proteins. Image published in: Bin-Nun N et al. (2014) Copyright © 2014. 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

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