Fig 1: Histone chaperone perturbation demonstrates their functional connectivity(A) Clustering analysis showing Euclidean distances between median normalized LFQ intensities (LFQM.N.) for proteins identified in H3.1 and H3.3 pull-downs by MS.(B) Bubble plot showing the changes in abundance of proteins identified in histone H3.1 and H3.3 pull-downs from extracts siRNA depleted of the chaperones ASF1A, ASF1B, NASP, and DAXX compared with control conditions (siCTRL). Colors represent Log2 ratios of median normalized LFQ intensities (siRNA/siCTRL), and radii represent p values.(A and B) Representative of n = 5 biological replicates, with median normalized LFQ intensities quantified on a protein or *peptide level by MS. Proteins are referred to by human UniProt protein identification code. See also Table S1.(C) Western blot of soluble extracts from cells expressing H3.1-FlagHA (left) and H3.3-FlagHA (right) siRNA depleted for ASF1A, ASF1B, DAXX, or NASP and compared with control knockdowns siCTRL. Representative of n = 5 biological replicates. See also Figure S4.
Fig 2: Histone chaperones collaborate through their histone-dependent associations(A) Network analysis of the histone-dependent interactors identified for ASF1a, ASF1b, NASP, DAXX, HJURP, DNAJC9, MCM2, and TONSL. Nodes and edges are colored based on their identifications in the different pull-downs.(B) Upset plot showing the overlaps between histone-dependent interactomes. Protein nodes, edges are colored based on their identifications in the different pull-downs, colored as in (A).(C) Clustering of histone-independent interactors using functional associations annotated in the STRING database and the MCL algorithm. Protein-protein functional associations are shown with a black line according to the string database, protein nodes colored as in (A).(A–C) Proteins are referred to by human UniProt protein identification code. Data generated from n = 4 biological replicates. See also Table S1 and Figure S1.
Fig 3: Histone chaperones directly interact with diverse cellular processes(A) The triple SILAC IP-MS strategy for mapping the histone chaperone network.(B) Network analysis of histone-independent interactors identified for ASF1a, ASF1b, NASP, DAXX, HJURP, and DNAJC9. Nodes and edges are colored based on their identification in the different pull-downs.(C) Upset plot showing the overlap of histone-independent interactomes, colored as in (B).(D) Clustering of histone-independent interactors using functional associations annotated in the STRING database and the MCL algorithm. Protein-protein functional associations are shown with a black line according to the string database, protein nodes colored as in (B). See also Figure S2A.(B–D) Proteins are referred to by human UniProt protein identification code. Data generated from n = 4 biological replicates. See also Table S1 and Figures S1 and S2.
Supplier Page from Abcam for Recombinant Human ASF1b protein (His tag C-Terminus)