FIGURE 4.

Histone H2A Phe-26 and Glu-57 residues contribute to the removal of H3K4me1 marks by Jhd2 demethylase in vivo. A, Western blot analysis of H3K4 methylation using nuclear extracts prepared from control wild-type, set1Δ, and sdc1Δ strains. B and C, Western blots showing H3K4me1, H3K4me2, and Jhd2 levels in crude nuclear extracts prepared from sdc1Δ strain expressing wild-type H2A or the H2A mutant and transformed with either an empty vector (pRS426) or a plasmid to overexpress Jhd2–9Myc. H3 serves as a loading control. *, a cross-reacting protein. C, blots were quantified by densitometry using ImageJ. Signals for H3K4me1 and H3K4me2 in each strain were normalized to the H3 signal. Graphs show -fold change in the normalized signals for H3K4me1 or H3K4me2 mark in various strains relative to the control sdc1Δ/H2AWT strain transformed with vector pRS426 (arbitrarily set as 1). Dotted line, decrease in H3K4me1 or H3K4me2 upon Jhd2 overexpression.