Figure 1: Quantitative ChaC for systems dissection of the functional G9a interactome or complexome formed within the chromatin of differentially inflamed macrophages. | Nature Communications

Figure 1: Quantitative ChaC for systems dissection of the functional G9a interactome or complexome formed within the chromatin of differentially inflamed macrophages.

From: A chromatin activity-based chemoproteomic approach reveals a transcriptional repressome for gene-specific silencing

Figure 1

(a) Workflow of the activity-based ChaC with immobilized UNC0638 coupled with AACT/SILAC quantitative proteomics. Equal amounts of nuclear extract from each set of BMDMs were incubated with Sepharose conjugated with UNC0638 or UNC2249 and the pull-down products were mixed and subjected to LC-MS/MS. (b) Immunoblot analysis of the G9a pulled down by UNC2249 from the BMDMs under different inflammatory conditions (N, NL and TL). Upper bands indicate the long isoform of G9a, while the lower bands are the shorter forms. β-Actin is the loading control. Same in figures below: all full blots are in Supplementary Fig. 14. (c) Scatter plot of the pull-down proteins with TL/N and NL/N ratio measured by AACT-based quantitative ChaC. The insets indicate previously known G9a interactors with their ratios to determine the threshold of NL- or TL-specific interactors. (d) Protein–protein interaction network of TL-specific G9a-interacting protein complexes, determined by STRING in high confidence (The confidence score is at 0.7).

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