FIG. 4.

The influence of H3K9ac on Bmh1 binding is less than that of H3K14ac. (A) (Top) Western blot (WB) of proteins bound to modified or unmodified (unmod) H3 and H4 peptides after incubation with yeast WCE containing three-HA-tagged Bmh1, washing in the presence of 350 mM NaCl, resolution by SDS-PAGE (4 to 20% gel), transferring to nitrocellulose, and probing with antibody against HA epitope (α-HA). (Bottom) Anti-HA Western blot of input WCE and anti-HA IP (4 to 12% gel). (B) Silver-stained gel of bound proteins from the same samples as in the experiment in panel A (4 to 20% gel). Lane M, molecular size standards. (C) Recombinant GST-tagged human 14-3-3ɛ bound to peptide resin after washing in the presence of 500 mM NaCl, resolution by SDS-PAGE (4 to 12% gel), and silver staining. (D) 14-3-3ɛ bound to peptide resin after washing in the presence of 150 mM NaCl, resolution by SDS-PAGE (4 to 12% gel), and silver staining. (E) Flowthrough (FT) fractions of unbound 14-3-3ɛ from the binding experiment in panel D (4 to 12% gel). The numbers to the left of the gels are the molecular sizes (in kilodaltons) of the proteins.