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Fig. 3. Co-injection of tsg mRNA rescues the tsg morphant phenotype. (A) Alignment of the target sequence for the tsg MO used in this study with the 5′ region of mRNAs for X. tropicalis, X. laevis and Oncorhynchus mykiss (rainbow trout). The translational start site is shown in green, mismatches between O. mykiss and the tsg MO are shown in pink, and mismatches between X. laevis and the tsg MO are shown in blue. (B) Embryos were injected at the two-cell stage with 20 ng tsg MO or 20 ng tsg MO plus 400 pg of O. mykiss tsg mRNA, then assayed at stage 26 for survival and proper blastopore closure. The experiment was repeated with the same result. (C) Embryos were injected with 400 pg O. mykiss tsg mRNA (b, f, j, n, r, v), 20 ng tsg MO (c, g, k, o, s, w), or both (d, h, l, p, t, x) and compared to uninjected sibling control embryos (a, e, i, m, q, u) at stage 11 (a–l), stage 19 (m–t), or stage 26 (u–x) for expression of the molecular markers indicated at left. The view for each marker is indicated at right. For animal and vegetal views, dorsal is to the top while for ventral and lateral views, anterior is to the left. |
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Fig. 7. BMP depletion rescues patterning defects in tsg morphants. Expression of region-specific markers were analyzed at stage 25 in uninjected embryos (A, E, I, M, Q, U, Y), and in embryos injected with 10 ng of tsg MO (B, F, J, N, R, V, Z), 20 ng each of BMP4 and BMP7 MOs (C, G, K, O, S, W, AA) or a combination of 10 ng tsg MO and 20 ng of BMP4 and BMP7 MOs (D, H, L, P, T, X, BB). Markers assayed included the midbrain hindbrain boundary marker en-2 (E H), the hindbrain marker krox-20 (I L), the spinal cord marker hoxb 9 (M P), the forebrain markers Xbf-1 (Q T) and eomesodermin (U X) and the ventral marker sizzled (YB). All views are lateral with anterior to the left and dorsal to the top. |
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Fig. 8. Chordin overexpression rescues tsg morphants. Expression of the ventral mesodermal marker sizzled was examined in stage 25 uninjected embryos (A), and in sibling embryos injected with intermediate doses of tsg MO (B), or an intermediate dose of tsg MO plus chicken chordin mRNA (C). Expression of the dorsal telencephalon marker eomesodermin was also compared among uninjected embryos (D), sibling embryos injected with an intermediate dose of tsg MO (E), and siblings injected with an intermediate dose of tsg MO and chicken chordin mRNA. All views are lateral with anterior to the left and dorsal to the top. |
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Supplementary Fig. S1. Rescue of the chordin phenotype by depletion of BMPs. Embryos were injected with chordin MO (B, F, J), BMP 4 and BMP 7 MOs (C, G, K) or the combination of chordin BMP4, and BMP7 MOs (D, H, L), and compared to uninjected sibling controls (A, E, I). Embryos were assayed by morphology (A), expression of eomesodermin (E), or sizzled (I). Embryos are at st. 258 and viewed laterally, anterior to the left, and dorsal to the top. |
