Xenbase Image ID: 137206

FIGURE 1:. Microinjected pri-miRNAs are not processed in oocyte nuclei but are in matured eggs. Radiolabeled pri-miRNA-21 (A) and pri-miRNA-29b-1 (B) were injected into individual nuclei of oocytes. To control for proper nuclear injection, the nuclear-retained U6 snRNA was coinjected. After incubation for up to 2 h, the individual oocytes were manually dissected into nuclei and cytoplasm. The corresponding nuclear and cytoplasmic RNAs of individual oocytes were extracted and separated on denaturing urea PAGE. An aliquot of the injected mixture is loaded (input), as well as markers corresponding to processed pre- and mature miRNAs (marker). pri-miRNAs injected into oocyte nuclei are processed poorly and get mostly degraded, whereas U6 snoRNA is stable (arrowheads mark weak, processed pre-miRNA bands). (C) pri-miRNA-29b-1 injected into eggs becomes processed into pre- and mature (insert) miRNAs. (D) RNA folding prediction of synthetic pri-miRNA-29b-1 used for injection. The processed band was cloned, and three independent clones were sequenced. Alignment with the predicted pre-miR29b-1 (gray) shows perfect alignment with the cloned pre-miRNA-29b-1 (top). This proves proper Drosha processing of microinjected pri-miRNA-29b-1 in Xenopus eggs. The processing sites are indicated by arrowheads. Image published in: Muggenhumer D et al. (2014) © 2014 Muggenhumer, Vesely, et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View

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