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. 2018 Jun 29;92(14):e00262-18. doi: 10.1128/JVI.00262-18

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FIG 5 — Chromatin localization of BKRF4 and BKRF4N. (A) CNE2Z cells were transiently transfected with plasmids expressing FLAG-BKRF4 or the indicated FLAG-tagged BKRF4 mutant. Log-phase cells were fixed and stained with anti-FLAG antibody and DAPI. Scale bar = 10 μm. (B and C) 293T cells were transiently transfected with plasmids expressing FLAG-BKRF4 (B) or the indicated FLAG-tagged BKRF4 mutant (C). Log-phase cells were lysed (total fraction in panel C) and separated into soluble and insoluble fractions. Chromatin-associated proteins in the insoluble fraction were then solubilized by DNase digestions to generate the chromatin-associated fraction (chromatin). Equal cell equivalents of each fraction were analyzed by Western blotting with anti-FLAG antibody. The same fractions were probed with antibodies against histone H4 and Nm23-H1 as markers of chromatin-associated and soluble proteins.