Xenbase Image ID: 137209

FIGURE 4:. Maturation of oocytes to eggs leads to an increase of Xenopus Drosha protein and poly A+ extension of Drosha mRNA. (A) qPCR of oocyte and egg cDNA shows threefold decrease in Drosha mRNA relative to Smn-2 mRNA. (B) Western blot of individual oocytes and eggs of two different frogs with a monoclonal anti-Drosha antibody. To verify that the antibody is recognizing the correct proteins, oocytes were also injected with mRNA encoding myc-xlDrosha. Drosha is well detected in eggs but barely visible in oocytes unless they were previously injected with RNA encoding myc-Drosha. (C) Scheme of poly(A) tail determination. cDNA was prepared with an anchoring primer. Depending on the length of the poly(A) tail, a short (top) or long (bottom) PCR product is obtained between a specific primer (P1) and an anchored primer (P2). (D) Whereas the poly(A) tail extends up to 90 nucleotides in oocytes (asterisk), an extension of up to 200 nucleotides can be observed in eggs (black bar). M, marker bands. The 60–base pair band in the poly(A) tail PCR lane most likely originates from priming of the oligo(dT) primer used for cDNA synthesis close to the poly(A) site. 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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