DEV028091 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. myog and myod MOs can specifically block translation of their mRNAs. (A,B) Western blot of protein extracts from control (A,B) and myog MO-1 (A) or myod MO (B), showing absence of specific bands at ∼37 kDa from the MO lanes. Myod immunohistochemical stain is also ablated (Hammond et al., 2007) #8381. (C) Immunohistochemistry with Myogenin antibody (brown nuclei) of myog MO-1-injected, myog MO-2-injected and control 15 ss embryos. Dorsal flatmounts of one side of anterior somites, anterior to top. (D) Confocal stacks of tail somites of 24 hpf control and myog MO-1-injected embryos after immunodetection of Myogenin (green nuclei). Lateral view, anterior to left. myog morphants lack the nuclear Myogenin reactivity of control embryos.

• Supplemental Figure S2 -

  Fig. S2. myf5^hu2022 mutation generates a likely null allele. (A) The genomic and protein structure of myf5 showing the nature of the hu2022 allele. (B) DNA sequencing traces from three individuals of each genotype. (C) Double in situ mRNA hybridization of myf5 (red) and the gene indicated (purple) in myf5^+/hu2022 incross embryos at 13-15 ss. Dorsal flatmounts, anterior to top. myf5^hu2022 mutants have low myf5 signal (arrowheads) compared with their siblings, but show no other change in mRNA expression.

• Supplemental Figure S3 -

  Fig. S3. mrf4^hu2041 mutation generates a likely null allele. (A) The hu2041 mutation in mrf4 and the resultant truncated protein. (B) Ectopic myogenesis in the head is induced by injection of wild-type mrf4 mRNA, but not by mrf4^hu2041 mutant mRNA. The full-length coding sequences of mrf4 and mrf4^hu2041 were inserted into the SacI-SalI sites of pβUT3 (gift from Dr Adam Rodaway, KCL). Capped mRNAs were transcribed from linearised plasmid using the T3 mMESSAGE mMACHINE Kit (Ambion), 100 pg/embryo injected at the 1- to 2-cell stage and embryos analysed by MyHC immunodetection at 24 hpf.

• Supplemental Table S1 - Adobe PDF