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. 2016 Aug 15;291(40):21171–21183. doi: 10.1074/jbc.M116.730218

FIGURE 1.

FIGURE 1.

Seven different human Importins bind the H3 and H4 tails. A, domain organization of human histones H3 and H4. B, pulldown binding assays of immobilized GST-H3 tail and GST-H4 tail (each 20 nmol; 250 nm) with a 500 nm concentration of each Importin (SDS-PAGE/Coomassie Blue). Relative densities of the gel bands from three separate experiments are plotted in histograms. One-way ANOVA tests were performed (H3 tail, p < 0.0001; H4 tail, p < 0.0001). Error bars represent S.D. C, Impα binding to immobilized GST-SV40 c-NLS, GST-H3 tail, and GST-H4 tail. Impα binds more tightly to the c-NLS than to the histone tails. D, binding assays of immobilized GST-H3 tail with Impβ, Kapβ2, Imp4, Imp5, Imp7, and Imp9 in the presence and absence of RanGTP (SDS-PAGE/Coomassie Blue). E, control experiments. Left panel, input samples of purified recombinant Importins (5 μl; 8 μm concentration of each protein; ∼10% of proteins used in B and in Figs. 2; 3C; 4, B–D; 5; and 6). Middle panel, Importin (∼4 μm) binding to GST protein (0.8 nmol; ∼2 μm) immobilized on glutathione-Sepharose. Right panel, Importin binding to immobilized GST-RanGTP. All six β-Importins bind RanGTP tightly.