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Dmrt2b is involved in slow muscle development through Hedgehog signaling. Analysis of slow muscle development by whole-mount immunostaining of 26 hpf embryos with antibodies labeling slow MyHC (F59) (A, C, E, G, I, K, M) and engrailed muscle pioneer cells (4d9) (B, D, F, H, J, L, N). Lateral view of embryos injected with Cont-MO (A, B), Dmrt2b-MO (C, D), embryos co-injected with Dmrt2b-MO+dmrt2b (E, F), Cont-MO+dnPKA (G, H), Dmrt2b-MO+dnPKA (I, J), Cont-MO+Su(fu)-MO (K, L), Dmrt2b-MO+Su(fu)-MO (M, N). White arrows in 4d9 panel indicate reduction of engrailed staining in the knockdown embryos compared to controls. All images show the somites over the yolk extension. Anterior is left in all images. (O) Quantitative analysis of slow muscle fiber number per somite in embryos at 26 hpf, injected as indicated. Data represent average±SD. *** indicates significance of p<0.0001 for Dmrt2b-MO vs. Cont-MO, Dmrt2b-MO+dmrt2b, Dmrt2b-MO+dnPKA and Dmrt2b-MO+Su(fu)-MO. n = 20 to 38 embryos per condition. (P) Quantitative analysis of muscle pioneer cells number per somite in embryos at 26 hpf, injected as indicated. Data represent average±SD. *** indicates significance of p<0.0001 for Dmrt2b-MO vs. Cont-MO, Dmrt2b-MO+dmrt2b and Dmrt2b-MO+dnPKA. n = 20 to 36 embryos per condition.
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