Skip to main content
. 2011 Jan 18;6(1):e15894. doi: 10.1371/journal.pone.0015894

Figure 5. PGVLL and ARKS motifs within the ADNP homeodomain are involved in HP1 chromoshadow domain interaction.

Figure 5

(A) Schematic representation of the ADNP-homeodomain (region aa 701–846), containing a K to R mutation in the ARKS and a V to E mutation in the PGVLL motif. (B) Fluorescence analysis of unfixed, living NIH3T3 cells inducibly expressing the indicated wild type (wt) and point mutant YFP-ADNP fusion proteins. DNA was stained with Hoechst 33342 dye. Bars, 5 µm. (C) Pulldown experiment with H3K9me3 peptides immobilized on magnetic beads using TNT-reticulocyte-extract expressed ADNP-FLAG (wt and point mutants) and recombinant wild type His6-HP1β. Western blot analysis of the reactions using the indicated antibodies is shown. (D) Pulldown experiment with H3K9me3 peptides immobilized on magnetic beads using wild type ADNP-FLAG protein expressed in TNT reticulocyte extract expressed together with recombinant wild type (wt) and W170A mutant (mt) His6-HP1β. Western blot analysis of the reactions using the indicated antibodies is shown.