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Transcription and translation in mouse blastocysts.

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posted on 20.02.2013, 04:19 authored by Telma C. Esteves, Olympia E. Psathaki, Martin J. Pfeiffer, Sebastian T. Balbach, Dagmar Zeuschner, Hiroshi Shitara, Hiromichi Yonekawa, Marcin Siatkowski, Georg Fuellen, Michele Boiani

mRNA levels were determined for pools of six ICSI and SCNT blastocysts (in duplicate), by quantitative RT-PCR, over two independent experiments. Technical replicates were determined three times for each sample, and results were normalized to Flag-Oct4 mRNA. (A) dCT values for transcripts in mtGFP-Tg SCNT, compared to ICSI. (B) dCT values for Gapdh and Actb mRNA, as well as protein levels (by immunocytochemistry), in wild-type SCNT relative to ICSI. Gapdh and Actb levels were significantly different, both for mRNA (t-test, p = 0,001 and p = 1,29E-08, respectively) and protein (t-test, p = 0.006 and p = 0.04, respectively). Box plot: fluorescence intensities distribution in the blastocyst; top and bottom lines: inter-quartile range; middle line: median; whiskers: range of variation limited to 1.5 times inter-quartile range.