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Fig. 3 | Genome Biology

Fig. 3

From: GATA3-dependent cellular reprogramming requires activation-domain dependent recruitment of a chromatin remodeler

Fig. 3

GATA3 induces nucleosome reorganization by targeting nucleosomal DNA. Metagene plots of normalized tag density of MNase-seq in G1 (a), G2 (b), and G3 (c) categories. Tag densities were normalized by the total counts in a +/- 1 kb window centered on the midpoint of GATA3 binding sites, then smoothed with a moving average (N  = 5). d The model of GATA3 bound nucleosome. GATA3 DBD and nucleosome core particle are represented in yellow and white, respectively. In the zoomed image, double-stranded DNA co-crystallized with GATA3 DBD is highlighted in cyan. The modified sequences are also indicated below. e Reconstituted nucleosome by salt dialysis method. The GATA consensus motif positive and negative DNAs were used to reconstitute mononucleosomes. f Purified recombinant GATA3 DBD. Purified proteins was analyzed by SDS-PAGE followed by staining with Coomassie. g Gel shift assay showing nucleosome binding of GATA3 DBD. The consensus motif containing nucleosome was used in lanes 1 to 4. The motif-mutated nucleosome was used in lanes 5 to 8. Nucleosomes (at a final concentration of 0.2 μM) were incubated with GATA3 DBD for 30 min at room temperature, and analyzed on native-polyacrylamide gel. The protein concentration of GATA3 DBD is as follows: 0 μM (lanes 1, 5), 0.5 μM (lanes 2, 6), 1 μM (lanes 3, 7), and 2 μM (lanes 4, 8)

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