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. 2018 Jan 16;115(5):E876–E885. doi: 10.1073/pnas.1717509115

Fig. 4.

Fig. 4.

CDCA7e is an essential stimulator of HELLS nucleosome remodeling activity. (A) Restriction enzyme accessibility nucleosome remodeling assay with HELLS–CBP and MBP–CDCA7e. The 601-positioned mononucleosomes with a PstI site engineered 15 bp into the nucleosome with 20 bp flanking DNA on each end were incubated with the indicated remodeling proteins and PstI. Productive nucleosome sliding exposes the PstI site, resulting in cleaved DNA (arrow). DNA was resolved on a 10% polyacrylamide gel and visualized with SYBR Gold. Representative of n = 2 independent experiments. (B) Native gel nucleosome remodeling assay with HELLS–CBP and MBP–CDCA7e. Center-positioned mononucleosomes (same as in A) were incubated with the indicated remodeling proteins. Reactions were stopped, resolved on a 5% polyacrylamide gel, and visualized with SYBR Gold. Sliding results in end-positioned nucleosomes, which migrate faster. Representative of n = 2 independent experiments.