Larger Image Figure 3. Long-Range Feedback for Expression of chordin and sizzled (A-C) qPCR analysis of transcriptional control for sizzled and bmp4 (A, ventral), myoD (B, lateral), and chordin, admp, and shh (C, dorsal) in the β-catenin-MO-injected embryo at stage 12 (n = 6; three experiments). The values in qPCR (per embryo) were normalized with the internal control (luciferase mRNA) included at the reaction (see Experimental Procedures). The expression levels of the wild-type embryo (Cont; no MO injection) was defined as 1. Increasing amounts of Noggin were injected into the blastocoel of the late blastula as in Figure 2A. Bars represent SD. (D-K) Expression of sizzled in embryos at stages 10 (D and H), 11 (E and I), 12 (F and J), and 13 (G and K). (D) through (G) show control embryos, and (H) through (K) show chordin-MO-injected embryos. (L-S) Expression of chordin in embryos at stages 10 (L and P), 11 (M and Q), 12 (N and R), and 13 (O and S). (L) through (O) show control embryos, and (P) through (S) show sizzled-MO-injected embryos. (T and U) Quantification of expression levels of sizzled (T) and chordin (U) by qPCR (n = 8; three experiments). Bars represent SD. (V) Schematic of the feedback loop involving Chordin, BMP, Sizzled, and Chordin proteinases. (W) Simplified view of the axis-wide long-range feedback by Chordin and Sizzled. Image published in: Inomata H et al. (2013) Copyright © 2013. Image reproduced with permission of the Publisher, Elsevier B. V.
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