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. 2008 Feb 25;28(9):2872–2883. doi: 10.1128/MCB.02181-07

FIG. 6.

FIG. 6.

The core region of NBD interacts with histone H3 in the nucleosome. (A) Site-directed cross-linking experiment of CP and AET-modified HMGN2 proteins. CP was incubated with or without AET-modified HMGN2-28C or with HMGN2-S24E,S28C and then irradiated with UV. The mixtures were separated by SDS-15% PAGE, followed by Coomassie blue staining. A high-molecular-weight cross-linked product is visible only in lane 3. CH, core histones. (B) Mobility shift assays of CP with HMGN2, HMGN2-S28C, and AET-modified HMGN2-S28C reveals that the modified protein binds to CP. (C) Two-dimensional PAGE analysis of the cross-linked product. Arrows indicate the direction of the first and second dimensions of electrophoresis. Note that the cross-linked product contained only HMGN2 and H3. (D) Western blot analysis of the cross-linked product. The cross-linked product was separated by SDS-PAGE and transferred to a polyvinylidene difluoride membrane, followed by Western blotting with anti-histone H3 or anti-HMGN2.