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Fig. 3 | BMC Developmental Biology

Fig. 3

From: Stable and bicistronic expression of two genes in somite- and lateral plate-derived tissues to study chick limb development

Fig. 3

Stable and bicistronic expression of Tomato and GFP fluorescent proteins in myogenic cells following chick limb somite electroporation with a muscle-specific regulatory element. a Forelimb somites of E2.5/HH15 chick embryos were electroporated with the PT2AL-p57/βactin-Tomato-T2A-GFP stable vector containing the Tomato-T2A-GFP cassette under the control of the p57MRE muscle-specific regulatory element. Six days after electroporation, at E8.5, forelimbs were collected for wholemount visualisation (b–d), immunostaining on transverse (e–j, n–p) or longitudinal (k–m) limb sections. b–d Both Tomato and GFP fluorescent proteins were expressed in forelimb muscles, visualised in whole mount embryos. e–g The Tomato and GFP expression was visualised in limb muscles on transverse limb sections. Higher magnifications of muscle transverse sections showed a general co-localisation of GFP+ nuclei with membrane Tomato (h-j, arrowheads). However, Tomato was not always associated with GFP due to the multinucleated statute of muscle fibres and membrane fluidity (h–j, arrows). Tomato and GFP expression was observed in MF20+ muscle fibres (k–m, arrowheads) and in MF20- cells (k–m, arrows). n–p Nuclear GFP or membrane Tomato expression driven by the p57 regulatory element was barely observed in Pax7+ muscle progenitors (n–p, arrowheads). Scale bars, (e–m) 50 μm (k–p)

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