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. 2020 Sep 24;22(5):4412–4422. doi: 10.3892/mmr.2020.11535

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Copyright: © Zhu et al.

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Figure 5. — Effect of NOTCH4_R CpG site 2 methylation on ETS1 binding affinity. (A) ETS1 overexpression in 293T cell lines. Null, cells transfected with a pcDNA3.1 empty vector; pcDNA3.1-ETS1, cells transfected with an pcDNA3.1-ETS1 vector. (B) EMSA was performed to evaluate the effect of CpG site 2 methylation on binding of the ETS1 transcription factor to the NOTCH4_R region. (C) EMSA and super-shift western blot analysis confirmed ETS1 protein binding to the NOTCH4_R region. Specific DNA/protein complexes are indicated by a red arrow. Met, methylated, Mut, mutant; EMSA, Electrophoretic mobility shift assay.