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. 2021 Jul 5;49(14):8024–8036. doi: 10.1093/nar/gkab588

Figure 2.

Figure 2.

LSH mediated transfer of mH2A-H2B dimer into the canonical nucleosomes. (A) Schematic graph to illustrate histone transfer assay in vitro. Canonical mono-nucleosomes (Mono_nuc) were immobilized on streptavidin (SA) beads, which were incubated with recombinant LSH, ATP and free reconstituted histone dimers in the transfer buffer. Incorporation of histone dimers in the nucleosome bound beads and free histone dimers in the supernatant fraction were evaluated by Western-blot analysis. (B) Immobilized reconstituted H2A containing mono-nucleosomes were incubated with recombinant LSH, ATP and free mH2A1_Flag–H2B dimer in the transfer buffer for 60 min at room temperature. Both beads and supernatant fractions were collected to detect the transfer of mH2A1–H2B dimer by LSH using western blotting. Histone H2B served as a control.