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. 2013 Apr 24;41(11):5769–5783. doi: 10.1093/nar/gkt314

Figure 2.

Figure 2.

Pseudo-octasomes split in half during low-salt dialysis. (A) Gel-shift image (native 7% PAGE), AFM image and height distribution of control H3 octamers assembled with 147-bp Widom 601 duplex DNA. (B) Same as (A) except using Cse4/α62 particles (native 6% PAGE). (C) Same as (A) except using H3/α62 particles (native 6% PAGE). In (A–C), DNA standards are included only as a rough guide, as migration of nucleosomal particles varies depending on gel concentration and running conditions. (D) AFM image and height distribution of H3 particles produced by gradient dialysis from 2 to 0.25 M, followed by step dialysis to 0.25 mM, conditions that increase aggregation. Bar in each image = 100 nm. DNA duplexes (147-bp 601) added to each sample provided an internal height standard. (E and F) Gel-shifted bands contain all four histones. Bands from native 6% gels were excised as indicated and loaded onto an SDS–PAGE gel to determine the histone composition of the particles. Particles were assembled using (E) H3 or (F) Cse4.