DEV00625 Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - Histone methylation patterns are disturbed in Ezh2^Del/Del embryos at the blastocyst stage. (A) Histone H3-me₁-K27 modification is strongest in the ICM of control embryos and only weak in cells of the trophectoderm (upper panel). By contrast, in parthenogenetic embryos from Ezh2-deficient oocytes (PG^–/– embryos) the H3-me₁-K27 staining in the ICM is markedly reduced (lower panel). The inserts show higher magnifications of TE cells. (B) Histone H3-me₂-K9 (upstate) specific modification is uniformly distributed in the ICM and the trophectoderm at the blastocyst stage (upper panel). Ezh2^Del/Del embryos do not show any detectable differences in staining pattern or intensity (lower panel).

• Supplemental Figure 2 - Peptide competition data for me₁-, me₂- and me₃-K27 antibodies. Mammalian core histones (H2A, H2B, H3 and H4) were resolved by SDS-PAGE and then blotted to nitrocellulose. Each probing was either performed in the absence or presence (1mg/ml) of specific peptide competitor as indicated. The bound antibodies were detected using the Amersham ECL kit according to the manufacturer’s recommendations.

• Supplemental Figure 3 - Dot-blot data on linear histone peptide for 2´-me₃-K27 antibodies characterisation. Differentially methylated peptides at different concentrations (as indicated) were blotted onto nitrocellulose and incubated with 2´-me₃-K27 antibodies. The bound antibodies were detected using the Amersham ECL kit according to the manufacturer’s recommendations.