Fig. 2

ADMP provides dorsal signal required in Spemann's organizer at the onset of gastrulation. ADMP knockdown was induced by ADMPMO injection (7 ng/embryo) and subjected to analysis of changes in gene expression. a, b qPCR analysis of the effects on dorsal and ventral gene expression during early gastrulation (stage 10–10.25) (a) and later during early/mid-gastrula (stage 10.25–10.5) (b). c–n Effect of ADMPMO injection on the domains of chordin (c–f), gsc (g–j), and ADMP (k–n) expression. Embryos injected with ADMPMO (4 ng) either dorsally or ventrally were fixed at the onset of gastrulation. f, j, n The arc of the domain of expression was measured, and the size of the domain of expression relative to control embryos is shown (%). o–r The specificity of the ADMPMO was further tested in rescue experiments. Embryos were injected dorsally (turquoise) with ADMPMO (3.4 ng/embryo) alone or with increasing amounts of RNA encoding the zebrafish (ZF) ADMP protein (10 or 20 pg RNA/embryo). o–q In situ hybridization analysis of the changes in the chordin expression domain in control (o), ADMPMO injected (p), and embryos co-injected with ADMPMO and zebrafish ADMP mRNA (20 pg/embryo) (q). r Summary of the quantitation of the changes in the size of the chordin expression domain. Chordin analysis: control, n = 81; ADMPMO dorsal, n = 48; ADMPMO ventral, n = 33, gsc analysis: control, n = 19; ADMPMO dorsal, n = 20; ADMPMO ventral, n = 15, ADMP analysis: control, n = 20; ADMPMO dorsal, n = 18; ADMPMO ventral, n = 14; and chordin rescue analysis: control, n = 25; ADMPMO, n = 25; ADMPMO + ZF ADMP 10, n = 19; ADMPMO + ZF ADMP 20, n = 25. p values were calculated compared to controls (a, b, f, j, n) except in the rescue experiments that were compared to the ADMPMO sample (r). Statistical test; Dunnett’s (ANOVA) multiple comparisons test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns not significant