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Figure 2


Fig. 2. (A) Mutation of Dph4. Top, wild-type (C57BL6/J) sequence. Bottom, mutant sequence. Large font, exon-4 sequence. Small font, intron-4 sequence. (B) RT-PCR analysis of mRNA from wild-type (WT), heterozygous (het) and homozygous mutant (mut) embryos. The wild-type allele produces a 246-bp fragment and the mutant allele produces one of 177 bp. The heterozygote contains both fragments. (C) Schematic illustrating the splicing pattern of the wild-type (upper) and mutant (lower) Dph4 gene. (D) Western blot to detect DPH4 protein in wild-type (WT), heterozygous (het) and homozygous mutant (mut) E12.5 embryos. The lower band is a non-specific reaction of the antiserum, which serves as a loading control. Tracks are from the same gel but lane order has been edited for clarity. (E) Native gel and western blot, probed to detect eEF2 protein in wild-type (WT), heterozygous (het) and homozygous mutant (mut) E12.5 embryos. Mutant protein has a –1 charge shift relative to that of heterozygotes and wild types, and therefore migrates faster. Tracks are from the same gel but lane order has been edited for clarity.





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