Extended Data Fig. 2.

Specificity of anti-H3K27 methylation antibodies used in this study.

a, Peptide sequences of H3.1, H3.3 and H3.10 surrounding K27 used for testing antibody specificity. Different forms with no methylation (me0), mono-methylation (me1), di-methylation (me2), and tri-methylation (me3) at K27 were used in all dot blots. b-c, Dot blots with serial dilutions of the different forms of histone H3 peptides described in a. The resulting membranes were probed with (b) α-H3K27me1 from Millipore #07-448 and (c) α-H3K27me3 from Millipore #07-449. Importantly, both α-H3K27 methylation antibodies cross react with the correct methylated form of H3.10 peptides, confirming that a lack of H3K27me3 detection in sperm chromatin (Fig. 1a) or on ectopically expressed H3.10-3xHA (Fig. 2e) is not due to poor antibody affinity. The experiment was repeated twice on two independent blots. e, Representative image of T1 htr4;htr5;htr8 plants expressing either untagged H3.10 under control of an H3.3 promoter (left) or endogenous H3.3 (right). Plants devoid of endogenous H3.3 and expressing only H3.10 and H3.1 (left) were developmentally stunted and completely sterile. This was evident in two independent experiments with individual htr4;htr5;htr8 T1 lines. Raw blots are provided in Source Data Extended data fig. 2.