Xenbase Image ID: 137567

Fig. 2. MO knockdown of BAF60c or misexpression of BAF60a or BAF60b variants inhibits myogenesis. (A) Electroporation of BAF60c-MO or control-MO into somites on one side, followed by in situ hybridization for myogenin (purple) and detection of the FITC-coupled MO (red). Whole-mount views and sections show that BAF60c-MO led to localized myogenin loss (arrows and arrowheads), whereas control-MO had no effect. (B) Percentage of embryos with an effect on myogenin expression after BAF60c-MO injection. (C) Western blot of pooled somites shows reduced BAF60c protein levels in BAF60c-MO electroporated somites, when compared with control-MO or to somites from the non-injected side (–MO). MO electroporations are mosaic and images shown in A give a spatial resolution, whereas the western blot in C averages what occurs in all cells across the somite. (D) Whole-mount double in situ hybridization and sections show that misexpression of BAF60a or BAF60b variants in somites leads to localized loss of myogenin expression. Myogenin is in purple (arrows and arrowheads) and GFP transcripts, which are expressed from a separate, co-injected plasmid, are in red. The ratio of BAF60 expression plasmid to GFP expression plasmid is 5:1. (E) Myogenin expression phenotypes observed after electroporation. (F) qPCR shows reduced myogenin expression in somites electroporated with BAF60a or BAF60b expression vectors when compared with GFP plasmid controls. Material from multiple embryos was pooled. Error bars indicate s.d.; *P<0.05 (t-test). my, myotome; nt, neural tube; nc, notochord. Scale bars: 50 μm. Image published in: Goljanek-Whysall K et al. (2014) © 2014. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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