Extended Data Fig. 1: Biochemical characterization of the Dot1L-H2BK34ub system.
From: H2B Lys34 Ubiquitination Induces Nucleosome Distortion to Stimulate Dot1L Activity
a-c, Reconstitution of the unmodified (WT), H2BK34ub and H2BK120ub octamer, the size exclusion chromatography (SEC) spectrograms are shown with 15% SDS-PAGE analysis, respectively. d, Purification of Dot1L(1-416). e, DEAE column-based ion-exchange chromatography of the H2BK34ub nucleosome. f, SYBR Gold-stained 4.5% native gels of fractions in e. peak1: canonical H2BK34ub octasome, peak2: H2BK34ub hexasome. g, Methyltransferase activities of Dot1L(1-416) on the unmodified (WT) nucleosome, H2BK34ub hexasome and H2BK34ub ocatsome. h, ESI-MS spectrum and deconvoluted ESI-MS (average isotope) spectrum of histone H3K79Nle with an observed mass of 15255.3 Da (calculated 15257.9 Da). i, j, Superose 6 Increase column-based SEC purification and 4-12% SDS-PAGE analysis of the Dot1L-H2BK34ub complex after glutaraldehyde crosslinking.
