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. 2021 Nov 8;10:e67952. doi: 10.7554/eLife.67952

Figure 6. Biochemical crosstalk between H4 sumoylation and H3 methylation in HEK293 cells.

Figure 6.

(A) Immunoprecipitation (IP) from HEK293 cells transfected with HA-Su3(ΔGG)-H4 (HSH4). Input (I) and eluate (E) lanes correspond to undigested bulk chromatin and eluted HA-tagged mononucleosomes containing HSH4. Antibodies targeting H3K4me1/2/3 were employed to detect the degree of H3K4 methylation in HA-tagged mononucleosomes. Total histone H3 in each sample was employed as an equal loading control. (B) FLAG-HA-Su3(ΔGG)-H4(Δ1–11) (suH4) protein expression in HEK293 cells and its localization to chromatin within 4 hr. Lanes: 1, Cytoplasmic fraction. 2, Nucleoplasmic fraction. 3, Chromatin fraction. (C) RPMK normalized chromatin immunoprecipitation (ChIP)-seq signals for SUMO-H4 (red, right y-axis) and H3K4me3 (blue, left y-axis) are plotted across length normalized gene bodies for 19,531 UniProt annotated protein coding genes plus 3 kb upstream of the transcription start site (TSS) and 3 kb downstream of the transcription end site (TES).

Figure 6—source data 1. Unedited intact gels and western blot membranes for all gels and western blot images shown in Figure 6.