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. 2018 Jul 10;1(4):e201800107. doi: 10.26508/lsa.201800107

Figure 4. H2A–H2B promotes FACT interaction with the tetrasome.

Figure 4.

(A) FACT (800 nM) was mixed with 60 nM or 120 nM H2A–H2B before addition of tetrasome (40 nM) assembled with 147-bp DNA. Double tetra: two (H3–H4)2 assembled on 147-bp DNA. (B) FACT (1.3 μM) was premixed with 650 nM H2A–H2B (labeled with Atto 647), and 150 nM tetrasome (labeled with Alexa 488) was added. Intermediate complexes (indicated by arrows) were enriched over M2 resin and analyzed by 5% PAGE. The marker (lane 1) is on the same gel, but intervening lanes were removed. (C) To determine if the two intermediate complexes (indicated with arrows) contain the same relative amount of H2A–H2B, FACT was mixed with an even larger excess of H2A–H2B (160 and 240 nM) before adding tetrasome.