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Fig. 3. col activation in FCM nuclei incorporated in the DA3 myofibre in Drosophila. (A-C) col transcription in wt DA3 muscle precursors, visualised by in situ hybridisation to col primary transcripts (red dots), immunostaining for Col (green) and nuclear staining (blue). (A'-C') Blue and red channels; (A''-C'') green channel. (A) Stage 14 embryo. The DA3 muscle precursor contains several nuclei; the two distalmost have already accumulated a high level of Col protein and activated col transcription. One central nucleus starts accumulating Col protein (lower inset) but does not yet transcribe col. Two other FCM have probably fused but not started to import Col protein (surrounded by a dashed line in A', upper inset). Another FCM has started engaging in the fusion process, (dashed notch in A'). (B) Stage 15 embryo. At this stage, each DA3 muscle nucleus contains high levels of Col protein and transcribes col. (C) Stage 16 embryo. All the DA3 muscle nuclei still contain high levels of Col protein but col transcription has almost completely ceased. (D,E) In situ hybridisation to col primary transcripts (red dots) in (D) wt and (E) col¹ mutant embryos (two segments are shown). A membrane-targeted form of GFP expressed under control of the col promoter (P9cg construct) allows the visualisation of the DA3 muscle (green). Note the complete absence of col transcription in col mutant embryos (E). The white arrowhead points to a dorsal md neuron expressing Col. Scale bar: 5 µm.

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