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. 2017 Apr 1;144(7):1273–1282. doi: 10.1242/dev.145722

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© 2017. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 2. — SET-4 is a histone H4K20 methyltransferase. (A) SET-4 promotes H4K20 methylation in vivo. Anti-SET-4, H4K20me2, H4K20me3 and H3 immunoblots of lysates from wild-type and set-4 mutant animals are shown. SET-4 protein is indicated by the arrowhead. Images are representative of four biological replicates. (B) Recombinant GST-SET-4 fusion protein methylates H4K20me1 in vitro. Percentage methylation of H4K20 peptide substrates by GST proteins fused to wild-type SET-4, mutant SET-4(S182F) or human SET-4 ortholog SUV420H2 is shown. Data represent mean values from three biological replicates. (C) MALDI spectra illustrating conversion of H4K20me1 to H4K20me2 by GST-SET-4. Monoisotopic masses (protonated) of peptide substrates are indicated with arrowheads. Spectra are representative of three independent experiments.