Extended Data Figure 6 |. Asymmetric mutant H2B nucleosomes are destabilized.
a, Denaturing gel analysis of purified wild-type, mutant, and H2A/H2B-CfaN-streptag dimers. b, Proof-of-concept assembly and purification of heterotypic nucleosomes with ubiquitinated H2B (H2Bub), which allows asymmetric nucleosomes to be distinguished by size. Numbers indicate molecular species shown in Fig. 3a. Hex., hexasomes. Het., heterotypic. c, Assembly and purification of heterotypic (het.) nucleosomes containing H2BE71K and H2BE76K. Assembly of wild-type (effectively homotypic) nucleosomes served as a control for the assembly process. 6/8 refers to hexasomes/octasomes. d, Raw data showing enhanced fluorescence for both homotypic and heterotypic destabilizing mutant MNs in the Nap1-mediated dimer exchange assay (n = 3 biologically independent samples). e, Raw data showing the thermal stability of homotypic and heterotypic mutant MNs. Data are shown in triplicate with a sigmoidal fit for the dimer melt. The Thalf is shown for each fit as a dotted vertical line and reported as the mean ± SD (n = 3).