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Figure 2


Fig. 2. Genotype distribution and SoxE gene expression in mouse embryos in which Sox10 is replaced by Sox8. (A) Embryos from crosses of Sox10+/Sox8ki males and females were sorted according to their genotype (wt, +/ki, ki/ki) as indicated below the bars. Embryos with an age between 10.5 and 12.5 dpc (white bars, n=150) were separately grouped from those between 14.5 and 18.5 dpc (black bars, n=111). (B) Comparison of expression levels for Sox10 in wild-type embryos at 10.5 and 12.5 dpc (white bars) and for Sox8 specifically expressed from the Sox10 locus in age-matched Sox10Sox8ki/Sox8ki embryos (black bars) with primers recognizing a common sequence in both transcripts using quantitative Lightcycler-RT-PCR. Transcript levels in each sample were normalized to ß-actin. After normalization, transcript levels in the wild type were arbitrarily set to 1. Experiments were repeated twice with material from two independently obtained embryos for each genotype and age. (C) Determination of the relative contribution of the wild-type allele to overall expression from the Sox10 locus in the indicated tissues of Sox10+/Sox8ki embryos at 13.5 dpc and 18.5 dpc. Values were obtained by comparing transcript levels from the wild-type allele to the ones from both alleles by quantitative Lightcycler-RT-PCR. Transcript levels were normalized to rpl8 and to corresponding transcript levels in tissues from age-matched wild-type embryos. sc, spinal cord; sn, sciatic nerve; st, stomach. (D) Western blot of nuclear extracts prepared from brains of 18.5 dpc wild-type (wt) and Sox10Sox8ki/Sox8ki (ki/ki) embryos with antibodies directed against SoxE proteins (Sox8/Sox10), Sox8 and RNA polymerase II (RNA Pol II). RNA polymerase II served as loading control. (E) Immunohistological analysis of transverse sections from wild-type and Sox10Sox8ki/Sox8ki embryos at 12.5 dpc using antibodies against Sox8 and Sox10. Additionally, EGFP autofluorescence was documented in both genotypes. Sox8 expression in the Sox10Sox8ki/Sox8ki embryos is essentially a composite of Sox8 and Sox10 expression in the wild type.





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