Expression of SYCE2 inhibits the interaction of HP1α with H3K9me3.

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Expression of SYCE2 inhibits the interaction of HP1α with H3K9me3. Expression of SYCE2 inhibits the interaction of HP1α with H3K9me3. (A, B) Coimmunoprecipitation of HP1α and H3K9me3 from lysates of mock cells and RPE cells stably expressing FLAG-SYCE2 (A) or from MCF7 cells transfected with a nontargeting siRNA control or SYCE2-targeting siRNA (B). Protein complexes were immunoprecipitated using the anti-HP1α antibody or normal goat IgG as a negative control and visualized by Western blotting using the anti-H3K9me3 or the anti-HP1α antibody. (C) Protein–protein binding assay using recombinant proteins for FLAG-SYCE2, HP1α, and H3K9me3. The HP1α recombinant protein was incubated with the indicated amounts of the FLAG-tagged SYCE2 recombinant protein for 30 min, followed by addition of the indicated amounts of the H3K9me3 recombinant protein for 1 more hour. The H3K9me3 protein bound to HP1α was detected by immunoprecipitations using the anti-HP1α antibody or normal IgG as a negative control and subsequent Western blot analysis using the anti-H3K9me3 antibody. Noriko Hosoya et al. LSA 2018;1:e201800021 © 2018 Hosoya et al.