XB-IMG-83699
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Figure S2.
Distinct Long-Range Effects of Chordin and Noggin on pSmad1 Accumulation, Related to Figure 1
(A-F) Immunostaining of phospho-Smad 1 (pSmad1) in the midgastrula embryo. Cryostat sections across the dorsal-to-ventral marginal zone were made parallel to the plane of the blastopore. (A') through (F') show high-magnification images of the insets. (A) and (A') show the control embryo. Strong pSmad accumulation was preferentially seen in the ventral-most region. (B) and (B') show the β-CN-MO embryo. Strong pSmad accumulation was observed throughout the embryo. (C) and (C') show β-CN-MO embryo with single-blastomere injection of noggin (25 pg: venus expression as tracer) at the 8-cell stage. No substantial pSmad accumulation was seen on either the injection or noninjection side, indicating that the effect of noggin spread to the opposite side. (D) and (D') show β-CN-MO embryo with single-blastomere injection of chordin (25 pg: venus expression as tracer) at the 8-cell stage. Substantial pSmad accumulation was observed on the side opposite to the injection region, indicating that the influence of chordin was distance-sensitive. (E) and (E') show β-CN-MO + sizzled-MO embryo with single-blastomere injection of chordin. The area of pSmad accumulation expanded toward the chordin-injection domain. (F) and (F') show β-CN-MO embryo with single-blastomere injection of chordin. In this case, sizzled mRNA (400 pg) was coinjected with β-CN-MO at the 2-cell stage. No substantial pSmad accumulation was seen even on the noninjection side, reminiscent of noggin injection in (C).
(G-J) Schematic summary of the differential effects of noggin and chordin in the reconstitution assay.
Image published in: Inomata H et al. (2013) Copyright © 2013. Image reproduced with permission of the Publisher, Elsevier B. V.
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