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. Author manuscript; available in PMC: 2018 Oct 13.
Published in final edited form as: J Mol Biol. 2017 Jul 17;429(20):2975–2995. doi: 10.1016/j.jmb.2017.06.024

Fig. 4.

Fig. 4

Interaction with MED25 relieves the DNA-binding autoinhibition from α-helix H4 of ETV4. (a) Electrophoretic mobility shift assay (EMSA) titrating MED25 ACID (500 – 1 nM) with a single concentration (50 nM) of ETV4 (top) or EHF (bottom). EMSA gels are representative examples of at least three replicates for each protein. (b) Using a single concentration of MED25 ACID (500 nM), ETV4 WT or indicated mutants were titrated (100 – 0.01 nM) to determine the KD of the ETV4-DNA interaction in the presence or absence of MED25. Filled circles represent a single experiment and the horizontal lines represent the mean and the standard deviation. “*” Indicates p < 0.05. The KD values for the ETV4-DNA interaction were (mean ± standard deviation): WT (+ MED25) = 2.1 ± 0.5 nM; WT (− MED25) = 3.4 ± 0.8 nM; Phe428Ala/Phe432Ala (+MED25) = 3.4 ± 1.0 nM; Phe428Ala/Phe432Ala (− MED25) = 3.7 ± 0.8 nM; Leu430Ala (+ MED25) = 1.9 ± 0.5 nM; Leu430Ala (− MED25) = 1.9 ± 0.4 nM.