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Fig. S3. Effect of the double knock down of Dmrt4 and Dmrt5 on Ebf2 expresion. (A) Frontal view of tadpole embryos injected unilaterally with the indicated morpholinos (10 ng for Dmrt4-MO and Dmrt5-MO, 20 ng for MOc) and hybridized with Ebf2. LacZ mRNA was coinjected as a tracer. Changes in the expression of Ebf2 were scored in individual embryos by comparing the injected and uninjected sides of embryos. Embryos were classified into three phenotypes: no changes, weak reduction and strong reduction or absence of expression. Left and right panels show examples of embryos showing weak and strong reduction of Ebf2, respectively. (B) RT-qPCR analysis of Ebf2 expression in animal caps (stage 14) derived from embryos injected with Noggin (100 pg), MOc (10 ng), Dmrt4-MO (10 ng) and Dmrt5-MO (10 ng), as indicated. Expression levels were normalized to GAPDH and compared to the level observed in Noggin injected caps, which was set to 1. Note that in neuralized animal caps and in embryos, the severity of the phenotype observed in double knockdown is weakly increased compared to the phenotype observed in single knockdown. |
