(A) I-TASSER prediction of ELF4 structure showing the backbone with a cluster predicted to form a binding pocket encircled and DNA binding residues with red arrow pointing at aa 187. (B) UCSF Chimera software analysis from predicted models of hydrogen bonds (green lines) with listed aa residue interactions of ELF4^T187 (blue, left) and ELF4^N187 (pink, right). (C) LIGPLOT software analysis from predicted models of Van der Waals interactions (pink) and hydrogen bonds, showing predicted interactions with aa within the same chain or proximal (across the black line) based on protein folding for ELF4^T187 and ELF4^N187. (D) Relative fluorescence units of luciferase promoter reporter (PRF1 and MDM2) for ELF4 T187N overexpression normalized to the WT control. (E) Independent repeats of ChIP quantitative PCR for overexpressed WT and T187N ELF4 bound to promoters (NT, nontransfected). (F) Salt-titrated extraction assay of chromatin-bound ELF4. Western blot run on a gel including the total cell lysate and the cytoplasmic and nuclear soluble fractions and a second gel including the chromatin-bound salt extraction. Showing the proteins detached with the increasing concentrations of salt from the chromatin-bound fraction after sequential extraction of the cytoplasmic, nuclear soluble, and chromatin-bound fractions (top, representative blot) with red lines indicating the significant difference in extraction between the WT and T187N ELF4 at lower salt concentrations. ELF4 expression normalized to loading control and subsequently to the wild-type control (bottom) across 3 independent experiments. The data represent mean ± SEM of ≥3 independent experiments. Western blot is representative of experiment repeated at least 3 times; *P < 0.05, **P < 0.01, ***P < 0.001, 2-tailed paired Student’s t test and Wilcoxon matched pairs signed-rank test.