Fig. 3

Analysis of Tg(Hsa.CTGF:nlsmCherry)^ia49 reporter activation. (A–A”) In vivo confocal images of Hsa.CTGF:nlsmCherry fluorescence at 20 hpf. (B) Confocal sagittal section of the eye at 24 hpf. (C) Lens of a Tg(Hsa.CTGF:nlsmCherry)^ia49 adult fish. (D) Confocal Z-stack projection at 48 hpf showing the transgene expression in the rhombencephalon (r) and the midbrain-hindbrain boundary (MHB) regions. (E) Confocal Z-stack projection of the trunk at 48 hpf. (F–G) Sections of the Z-stack projection in (E) are shown to highlight reporter activation in the somatic muscle (F), in the floorplate, the neural tube and the notochord (G). A strong signal is detected in a row of nuclei located between the neural tube and the notochord and representing the floorplate. The neural tube is only weakly positive. Below the floorplate, the notochord is expressing mCherry in the nuclei located just dorsally and ventrally with respect to the dark stripe of the notochord cells vacuoles. (H) Reporter fluorescence is visible in the epidermis along the whole embryo; here in a confocal Z-stack projection of the terminal portion of the tail at 48 hpf. (I–J) Confocal Z-stack projections of the rostral region of a 72 hpf reporter embryo. In the inset, a dorsal view highlighting the mCherry expression in the pectoral fins is depicted. (K) Confocal sagittal section of the heart region of a Tg(Hsa.CTGF:nlsmCherry)^ia49/Tg(myl7:GFP) double transgenic embryo at 22 hpf. (K’–K”’) Inset of K: mCherry channel (K’), GFP channel (K”), merge (K”’). (L) Confocal sagittal section of the heart at 48 hpf. (L’–L”’) Inset of L: mCherry channel (L’), GFP channel (L”), merge (L”’). (M–M’) Heart of a Tg(Hsa.CTGF:nlsmCherry)^ia49 adult fish. (N,O) Confocal Z-stack projections at 6 days post fertilization (dpf), showing the strong transgene expression in the whole intestine. (N’) Inset of N, single confocal sagittal section, bright field and mCherry merge, focusing on the reporter expression in the anterior intestine. (O’) Inset of O, single confocal sagittal section, bright field and mCherry merge, focusing on the reporter expression in the mid-posterior intestine. (P) Confocal sagittal section of the mid-posterior intestine of a 6 dpf Tg(Hsa.CTGF:nlsmCherry)^ia49/Tg(gut:GFP)^s854 double transgenic larva. (P’) Merge with bright field. r: rhombencephalon; MHB: midbrain-hindbrain boundary; v- ventricle; a: atrium; b.a.: bulbus arteriosus; s.b.:swim bladder; a.i.: anterior intestine; p.i.: posterior intestine. Scale bar: 100 μm.