Xenbase Image ID: 152330

Fig. 5. Morpholino oligonucleotides lipofection in the retina. (A) Retinal lipofection of GFP Mo interferes with GFP translation. Embryos were lipofected at stage 18 with GFP and a control Mo plus CD2, or with GFP and GFP Mo plus CD2. In stage 41 embryos, the intensity of GFP fluorescence in retinal cells was analysed. We used a filter to decrease the fluorescence light of the microscope so that low GFP fluorescent cells are below the detectable threshold. We then counted the number of lipofected retinas (CD2 positive) that also contain GFP-positive cells. While 79% (n=43 retinas) of CD2-positive retinas also contained GFP-positive cells when lipofected with a control Mo, only 19% (n=42 retinas) contained GFP-positive cells when lipofected with GFP Mo. (B) Xseb4R Mo block the translation of Xseb4R mRNA. Synthetic Xseb4R mRNA (containing the complementary sequence of Mo1) was injected into two-cell stage embryos together with Xseb4R Mo1 or control Mo. Stage 10 embryo lysates were then analysed by western blotting with a polyclonal anti-XSEB4R antibody and a monoclonal anti-αtubulin antibody (control). In the presence of control Mo, the anti-XSEB4R antibody recognises the 22 kD XSEB4 protein. The presence of Xseb4R Mo1 specifically abolishes Xseb4R translation. (C) Xseb4R loss-of-function delays differentiation of retinal cells. The proportion of retinal cell types observed in retinas co-lipofected with GFP plus a control Mo, GFP plus Xseb4R Mo1, or GFP plus Xseb4R Mo2 was determined. Both Mo give the same results. The statistical analysis was performed using the Student's t-test. *, P<0.05; **, P<0.001; ***, P<0.0001. Image published in: Boy S et al. (2004) Copyright © 2004. Image reproduced with permission of the publisher 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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