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. 2010 May 24;107(23):10454–10459. doi: 10.1073/pnas.1003064107

Fig. 1.

Fig. 1.

Instability of the H3T nucleosome. (A) H3T nucleosomes, reconstituted using 1.2 mg/mL total histones and 0.7 mg/mL DNA, were analyzed by nondenaturing 6% PAGE. Lane 1 indicates naked DNA. Lanes 2 and 3 indicate the H3T nucleosomes before and after a 55 °C incubation, respectively. DNA was visualized by ethidium bromide staining. Asterisks indicate bands corresponding to nonnucleosomal DNA–histone complexes. (B) The H3T and H3.1 nucleosomes were purified using a Prepcell apparatus, and were analyzed by nondenaturing 6% PAGE with ethidium bromide staining. (C) Histone compositions of the purified H3T and H3.1 nucleosomes were analyzed by 18% SDS-PAGE with Coomassie brilliant blue staining. (D) Salt titration. The nucleosomes were incubated in the presence of 0.4 M (lanes 1 and 5), 0.6 M (lanes 2 and 6), 0.7 M (lanes 3 and 7), and 0.8 M NaCl (lanes 4 and 8) at 42 °C for 2 h. The samples were analyzed by nondenaturing 6% PAGE with ethidium bromide staining. Lanes 1–4 and 5–8 indicate experiments with H3.1 and H3T nucleosomes, respectively. Bands corresponding to nucleosome monomers and nucleosome-nucleosome aggregates are indicated. Asterisks represent bands corresponding to nonnucleosomal DNA-histone complexes.