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Experiment details for myf5

Green YS and Vetter ML (2011) Assay

EBF proteins participate in transcriptional regulation of Xenopus muscle development.

Gene Clone Species Stages Anatomy
myf5.L laevis NF stage 14 to NF stage 15 presomitic mesoderm , paraxial mesoderm

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  Fig. 5. EBF2 and EBF3 are sufficient for muscle target gene expression hGR-XEBF2 or hGR-XEBF3 mRNA were injected into one cell of two-cell stage embryos, followed by DEX treatment from the late gastrula stage (stage 11/11.5) to the neurula stage (stage 14/15). hGR mRNA was injected in control embryos. β-gal mRNA was coinjected as a marker of the injected side. In all panels the right side is the injected side, showing the blue color of X-gal staining. The (purple) expression levels of myod (B and C), m-cadherin (E and F), seb-4 (H and I), and myf5 (K and L) are strongly upregulated by EBF2 and EBF3 (brackets), while expression of hGR alone does not change the expression level of the target genes (A, D, G, and J). The expression of actin alpha (N) and tnnc1 (P) is downregulated by EBF3. All panels show dorsal views.

Gene Clone Species Stages Anatomy
myf5.L laevis NF stage 20 to NF stage 21 presomitic mesoderm , somite , paraxial mesoderm

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  Fig. 6. EBF2 and EBF3 are necessary for muscle target gene expression (A-L) Two vegetal cells of eight-cell stage embryos were injected with either XEBF2 MO and XEBF3 MO together, or control MO. β-gal mRNA was coinjected as a marker of the injected side. The expression level of target genes was examined at stage 20/21. In all panels the right side is the injected side, showing the blue color of X-gal staining. The (purple) expression levels of myod (B and C), m-cadherin (E and F), seb-4 (H and I), and myf5 (K and L) are downregulated by XEBF2 MO and XEBF3 MO together or by NLS-DN-EBF (brackets), while control MO does not change their expression levels (A, D, G, and J). (M, N) Two vegetal cells of eight-cell stage embryos were coinjected with XEBF2 MO, XEBF3 MO and hGR-XEBF2 mRNA, followed by DEX treatment from the late gastrula stage (stage 11/11.5) to the early tailbud stage (stage 20). The expression of m-cadherin is downregulated without EBF activity (M), but expression was rescued in the presence of EBF activity (N). All panels show dorsal views.

Gene Clone Species Stages Anatomy
myf5.L laevis NF stage 35 and 36 to NF stage 37 and 38 somite , musculature of face , hypaxial muscle , jaw muscle , periocular region

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  Fig. 4. Expression patterns of muscle target genes In stage 35–37 embryos myod, m-cadherin, actin alpha, seb-4, tnnc1 and myf5 are all expressed in skeletal muscle including somites (black arrows), migrating hypaxial muscle anlagen (black arrowheads) and jaw muscle (yellow arrows). myod, m-cadherin, actin alpha, seb-4, and myf5 (A-D and F) are expressed in the somites, migrating hypaxial muscle anlagen and jaw muscle, and these expression patterns overlap with those of ebf2 and ebf3 (Figure 2). m-cadherin (B) is weakly expressed in a central band in somites, with expression throughout the somite. myf5 (F) expression in somites is weaker than other genes at this stage, and is expressed at the leading edge of migrating hypaxial muscle. tnnc1 (E) is expressed in the somites. actin alpha, seb-4, and tnnc1 are expressed in the heart (yellow arrowheads). All embryos show lateral views.