Xenbase Image ID: 77341

Figure 2. miR-24a knockdown results in a reduction in eye size. (A–F′) Hematoxylin and eosin staining of 12-μm sections in embryos injected with 20 ng of 24aMO in one cell at the two-cell stage, injected half on the right. The knockdown of miR-24a causes a reduction in eye size that does not occur until after stage 26 (A), first becoming prevalent at stage 28 (B). The reduction in eye size becomes obvious in stage 31 (C), stage 34 (D), stage 40 (E), and stage 45 (F,F′) embryos (see also Supplemental Fig. 1). (G) The size of the eye was measured at stage 40 and the ratio of the injected eye to the uninjected eye in each embryo was used to classify the severity of phenotypes. miR-24a knockdown caused many embryos to have a smaller eye, an effect that was rescuable by coinjection of miR-24a duplex RNA, but not of miR-133b duplex RNA. A mismatch morpholino also had no effect on eye size ratio (n = 121). (H) 24aMO functionally represses miR-24a. A GFP construct with two miR-24a recognition elements (24aMRE) when injected alone strongly fluoresces, but shows significantly lower levels of fluorescence when coinjected with duplex miR-24a RNA (n = 9; error bars are the standard error of the mean [SEM]). This effect is dependent on the interaction between miR-24a and the miR-24a recognition elements, because mutating either abrogates the effect. 24aMO is able to rescue fluorescence of 24aMRE by blocking the function of miR-24a when coinjected. Similar results were obtained with multiple experiments. RFP without miR-24a recognition elements was always coinjected, so that a measure of GFP repression was expressed as the relative fluorescence of GFP/RFP. Image published in: Walker JC and Harland RM (2009) Copyright © 2009. Image reproduced on with permission of the Publisher, Cold Spring Harbor Laboratory Press. This is an Open Access article. Larger Image Printer Friendly View

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