A H3.3-containing nucleosome immunoprecipitation followed by mass spectrometry identifies H3.3-interacting proteins. (A) Workflow for the identification of H3.3-nucleosome interacting proteins using an immunoprecipitation (IP)/mass spectrometry approach. (B) Gene Ontology analysis applied to the list of proteins coprecipitated with HA/FLAG-H3.3 and identified by mass spectrometry. (C) Western blots with lysate obtained from MEFs expressing wild-type and mutant versions of HA/FLAG-H3.3. Antibodies specific to histone H3 lysine modifications were used for immunoblotting and validation of lysine conversion to arginine. (D) Spectral counts associated with NuRD subunits obtained from HA/FLAG-H3.3 and HA/FLAG-H3.3K4R IPs.
