Transcription and translation in mouse blastocysts.
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.