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. 2007 Dec 13;27(1):168–178. doi: 10.1038/sj.emboj.7601960

Figure 3.

Figure 3

Insertion of poly(A) cassette affects imprinted expression of Cdkn1c in only a subset of embryonic tissues. (A) SNuPE analysis of Cdkn1c in e15.5 placenta and embryonic tissues from e15.5 day conceptuses derived from crosses of female SD7 mice and male mice heterozygous for the poly(A) cassettes. Exclusive expression of the maternal allele (T) is observed in all tissues examined from wt and Ap-mutant conceptuses. Imprinted expression is lost (indicated by incorporation dCTP into cDNA) in placenta, gut, heart, and brain from pA conceptuses while primarily maternal-specific expression is retained in liver, kidney, lung, and MEFs. (B) RFLP-RT–PCR analysis of Cdkn1c, Slc22a18, and Kcnq1 in multiple tissues from pA truncation mutants and wt littermates. Hot-stop RT–PCR products were digested by an allele-specific restriction enzyme (Supplementary Table S1) and separated by 10% PAGE. (C) Three independent RFLP-RT–PCR assays were performed for each of five tissues from two pA truncation-mutant embryos, two wt embryos, and one KvDMR1 deletion embryo for Slc22a18, Cdkn1c, and Kcnq1. Maternal and paternal bands were quantitated using the PhosphorImager. The histograms show the expression (average of 3–6 determinations plus STDEV) of the ‘relaxed' paternal alleles in the two mutants relative to that of the maternal allele. Expression of paternal alleles in wt tissues was undetectable.