Xenbase Image ID: 173272

Fig. 2. Sox5 and Sox6 enhance muscle gene transcription in Myod1-induced myogenic transcription assay, acting as a transrepressor except for slow muscle-specific genes. (A) In situ hybridization experiments show ectopic expression of Des and Actc1 (arrowheads) in embryos injected unilaterally either with Myod1 synthetic mRNA alone (100 pg), or co-injected with Sox5, Sox6, Sox15, Sox8 or Sox9 (400 pg each) and fixed at neurula stage. (B) Embryos were injected unilaterally either with Myod1 synthetic mRNA (100 pg) alone or co-injected with Sox15 or dCSox15 (400 pg each), fixed at neurula stage and probed with Des and Actc1. (C) Embryos were injected unilaterally either with Myod1 synthetic mRNA (100 pg) alone or co-injected with Sox6-R or Sox6-A (400 pg each), fixed at neurula stage and probed with Des and Actc1. The arrowhead indicates ectopic expression. Probe is shown in the bottom right corner of each panel. (D) Western blot with anti-flag antibodies of late gastrula embryos injected bilaterally either with 400 pg of Sox5, Sox6, Sox5-R, Sox6-R, Sox5-A, or Sox6-A synthetic mRNAs. Molecular weight markers are indicated in kilodalton (kDa). (E) Expression of Des, Actc1, Ckm, MyhE3, Mylpf, Tnnc2, MyhE19, Myh7L, Myh7S, Myl2 and Tnnc1 determined by semi-quantitative RT-PCR from RNA extracted of animal cap explants. Embryos were bilaterally injected with Myod1 synthetic mRNA (100 pg) alone or co-injected with Sox5, Sox5-R, Sox5-A, Sox6, Sox6-R or Sox6-A (400 pg each). Number of PCR cycles was indicated on the left. Slow muscle-specific genes are underlined. (F) Bar graph showing percentage of embryos with no, faint or strong ectopic expression. Chi-squared test analysis for ectopic Des expression of different Sox constructs: Myod1 versus Sox5 (P value< 0,0001), Sox15 (P value< 0,0001), dCSox15 (P value< 0,0001), Sox8 non-significant (ns), Sox9 ns, for ectopic Actc1 expression of different Sox constructs: Myod1 versus Sox5(P value< 0,0001), Sox15 (P value=0,0021), dCSox15 (P value< 0,0001), Sox8 ns, Sox9 ns, for ectopic Des expression of different Sox6 constructs: Myod1 versus Sox6 (P value= 0,0140), Sox6-R (P value< 0,0001), Sox6-A (P value< 0,0001), for ectopic Actc1 expression of different Sox6 constructs: Myod1 versus Sox6 (P value< 0,0001), Sox6-R (P value< 0,0001), Sox6-A (P value= 0,0071). Numbers of injected embryos are indicated at the bottom of each bar. Image published in: Della Gaspera B et al. (2018) Copyright © 2018. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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