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. 2008 Oct 4;18(1):118–127. doi: 10.1093/hmg/ddn322

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© 2008 The Author(s)

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — TaqMan quantitative real-time PCR (qRT–PCR) measurements of Blcap_v1a and Blcap_v2a. Error bars indicate the 95% confidence interval. (A) Blcap_v1a and Blcap_v2a levels in MatDp(dist2) and PatDp(dist2) newborn (nb) brain relative to a matched non-UpDp control (T11H). Blcap_v1a was ∼2.2 times more abundant in MatDp(dist2) than in control nb brain, and achieved only ∼25% of the control expression level in PatDp(dist2) nb brain, which corresponds to a MatDp(dist2)/PatDp(dist2) expression ratio of ∼8.8. This reflects the preferential maternal expression of Blcap_v1a. Blcap_v2a was not present in MatDp(dist2) nb brain (no amplification), and ∼2-fold more abundant in PatDp(dist2) than in non-UpDp nb brain, consistent with paternal-only expression. (B) Left bar: amount of Blcap_v2a in PatDp(dist2) nb brain relative to Blcap_v1a in MatDp(dist2) nb brain. Right bar: amount of Blcap_v2a in non-UpDp nb brain relative to Blcap_v1a in the same sample. In both cases, Blcap_v2a reached only ∼1% of the Blcap_v1a transcript level, i.e. in nb brain, Blcap_v1a is ∼100-fold more abundant than Blcap_v2a. (C) Quantification of Blcap_v1a and Blcap_v2a levels in 8.5 dpc Dnmt3l^−/+ embryos relative to wild-type (WT) litter-mates. Expression of Blcap_v1a was reduced to ∼50% of WT level in Dnmt3l^−/+ embryos, while Blcap_v2a increased in abundance by ∼40%. This is consistent with the notion that the maternal allele-specific methylation of the Nnat promoter CGI is established during oogenesis and that methylation of this CGI positively regulates Blcap_v1a expression, while it inhibits Blcap_v2a expression.