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Figure S5. Noggin4 acts cell-non-autonomously when promoting the expression of anterior markers. (a) Control
embryo injected in dorsal blastomeres with EGFP mRNA mixed with FLD tracer at 8-cell stage and hybridised at the
midneurula stage with FoxG1 probe. This embryo is presented for comparison of width of the control FoxG1 expression band
with width of FoxG1 expression band in the embryo injected with Noggin4 mRNA and shown on b. (b-f) Midneurula embryos
injected with Noggin4 mRNA mixed with FLD tracer unilaterally, in one of the animal dorsal blastomeres, at 8-cell stage and
hybridised with probes to the indicated marker genes. Arrows indicate areas in which overexpression of the tested marker genes
does not match the localisation of FLD, thereby confirming cell-non-autonomous activity of Noggin4. (a’-f’) Fluorescent
images of the same embryos as on a–f. (a”–f”) Superimposed images of a–f and a’–f’. (g–g”). An expanded cement gland on
the non-injected side of the tail-bud stage embryo (right embryo), which was injected in the right animal dorsal and ventral
blastomeres at 8-cell stage, confirms cell-non-autonomous activity of Noggin4. |
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Figure 1: Properties of Noggin4. (a) Alignment of Xenopus laevis Noggins. Positions essential for Noggin1 binding to BMP1 are indicated by red arrows. HSPG binding motifs and cleavage sites are indicated by green and red dashed lines, respectively. (b) Whole-mount in situ hybridization of a midneurula embryo with dig-probe to Noggin4 mRNA. Dorsal view at the top, the right-frontal view at the bottom. (c) Effects of Noggin4 mRNA (80%, n = 120) and MO1 (85%, n = 130) injections on the development of head structures. Cg-cement gland. (d–s) In situ hybridization with dig-probes to FoxG1, Rx, Otx2 and Pax6 mRNA of midneurula embryos injected with GFP mRNA (control) (0%, n = 40; 0%, n = 50; 0%, n = 40; 0%, n = 30, abnormal phenotypes respectively), Noggin4 mRNA (85%, n = 30; 61%, n = 35; 81%, n = 40; 65%, n = 40 abnormal phenotypes respectively), Noggin4 MO1 (90%, n = 50; 86%, n = 40; 60%, n = 40; 60%, n = 40, abnormal phenotypes respectively) and misNoggin4 MO1 (control) (0%, n = 30; 0%, n = 30; 0%, n = 40; 0%, n = 30, abnormal phenotypes respectively). Red dashed line surrounds the presumptive rostral forebrain territory. Anterior view, dorsal to the top. |