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. Author manuscript; available in PMC: 2015 Jun 11.
Published in final edited form as: Nature. 2014 Sep 24;516(7530):267–271. doi: 10.1038/nature13736

Figure 2. CK2 phosphorylates Y57 in H2A.

Figure 2

(a) CK2α interacts preferentially with the H2A-Y57F mutant. Flag-tagged WT and H2AX-Y57F were expressed in 293T cells, immunoprecipitated and immunoblotted. (b) CK2 phosphorylates Y57 in H2A in nucleosomes in vitro. In vitro kinase assays were performed using recombinant GST-CK2α, and full-length or nucleosomal Flag-tagged H2AX purified from 293T cells, and were immunoblotted to examine tyrosine phosphorylation. (c, d) CK2 phosphorylates Y57 in H2A in vivo. (c) CK2α was knocked down in 293T cells, and nuclear extracts were immunoblotted. CTL indicates scrambled siRNA. (d) Nuclear extract from 293T cells treated with TBBz were immunoblotted. Data represent three independent experiments.