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Fig. 3 | Epigenetics & Chromatin

Fig. 3

From: Tri-methylation of ATF7IP by G9a/GLP recruits the chromodomain protein MPP8

Fig. 3

G9a/GLP binds preferentially to unmethylated ATF7IP and its K16R mutant. a, b Co-IP assay using anti-FLAG M2 beads and HEK293T cells co-transfected with G9a (a) or GLP (b) plus either 3xFLAG-tagged ATF7IP WT, Δ(1–16) mutant, or K16R mutant. c mESC lines expressing 3xFLAG-ATF7IP WT, K16R, or Δ(1–16) mutant were used for co-IP assay with anti-FLAG M2 beads. Co-IPed G9a, GLP, and SETDB1 were detected by western blot analysis. d, e In vitro methylation followed by in vitro binding assay. Recombinant proteins produced and purified from E. coli were incubated with SAM for the indicated times, and the reactants were then used for western blot analysis or GST-pull-down assay. Pulled-down ATF7IP (residues 1–364) recombinant protein was detected by western blot analysis. The interaction of ATF7IP with both G9a (d) and (e) was decreased in the incubation time-dependent manner

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