Figure 3

Expression of the endogenous E11 and E1 mRNAs in wide-type C57BL/6J mice during ontogeny. A. RT-PCR: The temporal expression of a 184 bp exons 1/2 (Exon 1/2) and a 178 bp exon 11 (Exon 11)transcripts during ontogeny of C57BL/6J mice was determined by a relative quantitative RT-PCR approach as described in the Methods section. RNA input was estimated by a parallel PCR with a pair of G3PDH primers. Agarose gel stained with ethidium bromide was photographed with FluorChem 8000 system. PCR cycles and extension time at 72°C: for Exon 1/2 primer set, 35 cycles and 20 sec; for Exon 11 primer set, 45 cycles and 20 sec; and for G3PDH primer set, 25 cycles and 2 min. B. Relative quantification: Relative band intensities from the gel were quantified with AlphaEase FC software, and converted to amole concentration based upon the linear regression equations from the linear phase of each saturation curve (see Fig. 8) using Prism 4.0. The concentration was normalized with the G3PDH concentration. Right Y axis represents the concentration of Exon 11 cDNA, and left Y axis, the concentration of Exon 1/2 cDNA. Bars represent the mean ± S.E. of the concentration from three independent experiments.