Evi1 represses PTEN expression via its first zinc finger domain. (A) Schematic representation of Evi1 and its mutants. (B) Relative luciferase activity of Evi1 and its mutants on PTEN4_wt promoter (n = 4) together with immunoblot of assayed Jurkat cells transfected with Flag-tagged wild-type Evi1, its mutants, and mock. Error bars indicate SD; *P < .05, **P < .01, and ***P < .001. (C) PTEN mRNA expression in Flag-tagged Evi1-, its mutants-, or mock-transduced BM cells together with protein expression of Evi1 and its mutants (n = 3). Error bars indicate SD; *P < .05, **P < .01, and ***P < .001. (D) Protein expression of Flag-tagged wild-type Evi1 and its mutants used in EMSAs. Western blot analysis was done with anti-Flag antibody. (E) EMSAs used mutants of human Evi1 and the probe set 3. (F) ChIP analysis for Flag-Evi1, Flag-Evi1_ΔZF1-7, or mock-expressing BM cells that used the indicated antibodies (n = 3). Error bars indicate SD; *P < .05, **P < .01, and ***P < .001. REP, repression domain; Cend, domain at the C-terminus.