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. 2009 Apr 15;23(8):951–962. doi: 10.1101/gad.1769209

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Copyright © 2009 by Cold Spring Harbor Laboratory Press

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Figure 5. — The RING finger domain of Not4 is required for H3 K4 trimethylation and Jhd2 protein stability. (A) Schematic of Not4 illustrating positions of RING and RRM domains. Numbers indicate amino acid positions. (B) Whole-cell lysates from yeast strains expressing the indicated constructs were analyzed for histone H3 K4 methylation by immunoblotting with H3 K4 mono-, di-, or trimethyl-specific antibodies. Histone H3 levels and expression levels of Not4-HA and Not4-HA deletion mutants were verified using α-H3 and α-HA antibodies. (C) Jhd2 expression was analyzed in a not4Δjhd2Δ strain expressing the indicated Not4 constructs with a α-Flag antibody. HA-tagged Not4 and Not4 deletion mutants are detected with a α-HA antibody. Histone H3 is used as a loading control.