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Figure 5 (A) Nanobody localization in the neural tube of a zebrafish embryo. The mRNA of the anti-mCherry mNphp3(201)-VHHLaM-2-eGFP nanobody was injected into nacre zebrafish embryos. Embryos were stained with an anti-acetylated tubulin antibody (magenta, ciliary marker), an anti-GFP antibody (green), and DAPI (blue). (B) See A. for spinal cord. (C) See A. for eye. (D) RFP (red) expression in the neural tube of Ubi:zebrabow (Pan et al., 2013) transgenic embryos. (E) RFP (red) expression in the neural tube of Ubi:zebrabow (Pan et al., 2013) transgenic embryos, injected with mRNA of the anti-mCherry mNphp3(201)-VHHLaM-2-eGFP nanobody. Scale bars: 20 μm, magnified view: 10 μm. Boxes indicate the position of the magnified views shown at the bottom right as inset (A-C) or as a separate panel next to the overview image (D, E). Arrows in different colors indicate the direction and the length of the shift of the respective fluorescence channel. The upper right panel in D and E shows the RFP channel only, the bottom right panel shows the magnified view. A: anterior, P: posterior, L: left, R: right, D: dorsal, V: ventral. All images were taken from fixed samples.