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Tlr4 activates NFκB to stimulate frzb-mediated restriction of dorsal organizer formation. a–c Inhibition of Tlr4 reduces frzb expression and enhances Wnt/β-catenin signaling and dorsal organizer formation. Embryos were injected with mKO2 (control) or Tlr4 DN mRNA. WISH for (a) dGFP in NFκB-tkP:dGFP-transgenic; frzb and axin2 in WT, (b) dharma and chordin in WT embryos at the indicated stage. Animal views. Box plots of the angle of marker genes show first and third quartile, median is represented by a line, whiskers indicate the minimum and maximum. Each dot represents one embryo. P-values from unpaired two-tailed t-tests are indicated. c Representative pictures of 27 hpf larvae, lateral views with anterior to the left. The strength of dorsalization was scored. d–f Forced activation of Tlr4 by injection of lipopolysaccharide (LPS) activates NFκB signaling and inhibits dorsal organizer formation. d Phenotypes of 27 hpf larvae injected with LPS, uninjected as control. The strength of ventralization was scored. Lateral views with anterior to the left. e Embryos were injected with LPS and treated with DMSO or TAK-242 or co-injected with Tlr4 DN, uninjected as control. DMSO and TAK-242 were treated from 3 hpf to dome stage. WISH for dGFP in NFκB-tkP:dGFP-transgenic; frzb in WT embryos at dome stage. f WISH for axin2, dharma and chordin in embryos injected with LPS, uninjected as control. Animal views. Scale bar = 200 μm. Source data are provided as a Source Data file.
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