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. Author manuscript; available in PMC: 2022 Feb 4.
Published in final edited form as: Cell. 2021 Jan 21;184(3):643–654.e13. doi: 10.1016/j.cell.2020.12.022

FIG. 3. Y518F disrupts cross-linking but not binding to oriP.

FIG. 3.

A) 293T cells expressing FLAG-EBNA1 WT or Y518F with oriP were assayed by ChIP assay under standard formaldehyde cross-linking conditions (Cross-linked) or without formaldehyde (Native) and assayed for ChIP with antibodies to IgG or Flag (EBNA1 WT or Y518F) or H3K4me2, as indicated. ChIP-DNA was assayed for oriP FR sequence by Taqman specific qPCR. student T-test, n=3., ns -not significant, * p value < 0.01 ** p value <0.001*** p value <0.0001. B) EMSA analysis of EBNA1 DBD WT, Y518A, Y518F, Y518E, H468A or N519A using DS probe with a single EBNA1 binding site. EC) Coomassie stain of SDS-PAGE for 200 ng purified EBNA1 DBD WT, Y518A, Y518F, Y518E, H468A or N519A. D) Quantification of EMSA and calculation of EC50 (nM) for each mutant.