Generation of mutant mice harboring the point mutation translated into EVI1 H752R and MDS1-EVI1 H942R in mice, corresponding to EVI1 H751R and MDS1-EVI1 H939R in humans. (A) The point mutation is located in the eighth zinc finger motifs of EVI1 and MDS1-EVI1. The sequence of the eighth zinc finger motif is given in a box. “C” and “H,” shaded in a gray, bind to a zinc ion. EVI1 H751 in humans, MDS1-EVI1 H939 in humans, EVI1 H752 in mice, and MDS1-EVI1 in mice are represented in red letters. (B) Overexpression of C-terminal FLAG-tagged wild-type or mutant hEVI1/mEVI1 in NIH3T3 cells. (C) Luciferase assay for assessing the effects of wild-type or mutant hEVI1/mEVI1 on AP-1 and TGF-β signaling. pCAGGS empty vector (mock) indicates the effectiveness of newborn calf serum or TGF-β1 stimulation. The relationship between the reactions against the stimulation between wild-type and mEVI1 H752R resembles that between wild-type and hEVI1 H751R. Representative data from 3 experiments performed in triplicates are shown. Values are the mean ± standard error of the mean (SEM) of 3 samples of the representative experiment. Underlined numbers denote the P value (2-tailed Welch t test). Threshold significance level, P = .05.