Figure 3.
Che-1 displaces HDAC proteins from histones. (A) Kms27 nuclear extracts were immunoprecipitated with anti–Che-1 polyclonal antibody and analyzed by western blot (WB) analysis with the indicated antibodies. (B) Coomassie blue stain of histone proteins incubated with purified GST-Che-1 fusion protein or control GST agarose beads. (C) Sequence alignment of Che-1 and HDAC class I proteins. Conserved amino acid residues are in bold and italic. (D) Nuclear extracts from Kms27 cells transiently transfected with the indicated expression vectors were immunoprecipitated with anti-Myc 9e10 monoclonal antibody and analyzed by WB with the indicated antibodies. (E) Nuclear extracts from inducible ind-shChe-1 Kms27 cells treated or not with doxycycline (Dox) were immunoprecipitated with anti-HDAC1 polyclonal antibody and analyzed by WB with the indicated antibodies. (F) Coomassie blue stain of histone proteins incubated with purified GST-HDAC1 fusion protein or control GST agarose beads in the presence or absence of the indicated peptides. (G) Nuclear extracts from Kms27 cells transiently transfected with the indicated expression vectors were immunoprecipitated with anti-histone H3 antibody and analyzed by WB with the indicated antibodies. (H) Enrichment density of HDAC1 ChIP-seq significant peaks identified in siRNA control (siControl) (blue) and siRNA Che-1 (siChe-1) (yellow) samples. Enrichment density was calculated on the relative size of each peak (start to end of each fragment) extended 500 bp upstream and downstream. Boxplot shows the peak sizes. (I) HDAC1 ChIP-seq signal in siControl (blue) and siChe-1 (yellow) samples at AATF and c-Myc loci. Signal enrichment scale on the y-axis of each site. (J) Kms27 cells transiently transfected with siControl or siChe-1 were subjected to ChIP-qRT using anti-HDAC1 antibody followed by qRT. Data are expressed as percentage of input. Data represent the mean ± standard deviation. P value was calculated by Student t test (n = 3, *P < .0018, **P = .00034). See also supplemental Figure 3.

Che-1 displaces HDAC proteins from histones. (A) Kms27 nuclear extracts were immunoprecipitated with anti–Che-1 polyclonal antibody and analyzed by western blot (WB) analysis with the indicated antibodies. (B) Coomassie blue stain of histone proteins incubated with purified GST-Che-1 fusion protein or control GST agarose beads. (C) Sequence alignment of Che-1 and HDAC class I proteins. Conserved amino acid residues are in bold and italic. (D) Nuclear extracts from Kms27 cells transiently transfected with the indicated expression vectors were immunoprecipitated with anti-Myc 9e10 monoclonal antibody and analyzed by WB with the indicated antibodies. (E) Nuclear extracts from inducible ind-shChe-1 Kms27 cells treated or not with doxycycline (Dox) were immunoprecipitated with anti-HDAC1 polyclonal antibody and analyzed by WB with the indicated antibodies. (F) Coomassie blue stain of histone proteins incubated with purified GST-HDAC1 fusion protein or control GST agarose beads in the presence or absence of the indicated peptides. (G) Nuclear extracts from Kms27 cells transiently transfected with the indicated expression vectors were immunoprecipitated with anti-histone H3 antibody and analyzed by WB with the indicated antibodies. (H) Enrichment density of HDAC1 ChIP-seq significant peaks identified in siRNA control (siControl) (blue) and siRNA Che-1 (siChe-1) (yellow) samples. Enrichment density was calculated on the relative size of each peak (start to end of each fragment) extended 500 bp upstream and downstream. Boxplot shows the peak sizes. (I) HDAC1 ChIP-seq signal in siControl (blue) and siChe-1 (yellow) samples at AATF and c-Myc loci. Signal enrichment scale on the y-axis of each site. (J) Kms27 cells transiently transfected with siControl or siChe-1 were subjected to ChIP-qRT using anti-HDAC1 antibody followed by qRT. Data are expressed as percentage of input. Data represent the mean ± standard deviation. P value was calculated by Student t test (n = 3, *P < .0018, **P = .00034). See also supplemental Figure 3.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal