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. Author manuscript; available in PMC: 2014 Dec 10.
Published in final edited form as: Nat Genet. 2013 Sep 29;45(11):1386–1391. doi: 10.1038/ng.2777

Figure 4.

Figure 4

NSD2 p.E1099K alteration leads to increased enzymatic activity and promotes transformation. (a) Biochemical characterization of the enzymatic activity of wild-type (WT) and p.E1099K-mutant NSD2. The catalytic domain of NSD2 (955–1365) was purified and assayed with nucleosome substrates. Enzymatic activity was assessed by quantifying the production of S-adenosyl-L-homocysteine (SAH) on the y axis. Error bars indicate the s.d. of triplicate measurements. (b) Western blot analysis of lysates from KMS11-TKO cells reconstituted with WT NSD2, p.E1099K-mutant NSD2, CDM NSD2 or both p.E1099K and CDM (p.E1099K-CDM) NSD2 or empty vector, analyzed with the indicated antibodies. Histone H3, loading control. (c) Quantification of soft-agar transformation (5% serum) of KMS11-TKO cells reconstituted with the indicated NSD2 variants. Error bars indicate the s.d. of triplicate samples (see also Supplementary Fig. 5). (d) Global chromatin profiling of KMS11-TKO engineered lines. Chromatin-signature profiling, clustering and color coding are as described in Figure 1. KMS11-TKO cells were reconstituted with the indicated NSD2 variants. The chromatin-signature profiling of KMS11-TKO reconstituted cells is shown with a representative panel of hematopoietic cell lines (see Supplementary Fig. 4 for complete data set and dendrogram shown).