Correlation between ACE and CDX2 and expression. (A) Endogenous ACE mRNA expression by RT-qPCR in K562 and Nalm6 cells transfected with the control vector pFlag-CMV2 (−) or with the CDX2-expressing plasmid pFlag-CDX2 (+). (n = 3). Western blots for CDX2 and actin as control are illustrated below the graphic. (B) Luciferase activity in K562 and Nalm6 cells cotransfected with the reporter plasmid pACE-Luc containing the ACE promoter, or with the mutated form pACEm-Luc, together with the control plasmid pFlag-CMV2, with pFlag-CDX2 and/or with pTBPspm3. Values are expressed relative to the luciferase activity measured in cells transfected with the control luciferase plasmid pGL3-basic and presented as mean plus or minus SEM (n = 3). (C) Chromatin immunoprecipitation (ChIP) with anti-CDX2 or anti-Flag antibodies (Ab) in K562 cells transfected with pFlag-CDX2 (+) or pFlag-CMV2 (−), followed by PCR amplification of the ACE promoter fragment overlapping the CDX2-binding element. Specificity of the chromatin immunoprecipitation was assessed using immunoglobulin G (IgG). (D) ACE and CDX2 mRNA levels measured by RT-qPCR in the BMMCs from AML patients (AML-BMMC) and from healthy individuals (Ctr-BMMC). (E) Pearson correlation coefficient (r) and P value (p) between CDX2 and ACE expressions measured by RT-qPCR on BMMCs of AML patients. ***P < .001, ****P < .0001.