STAT3 regulation of CD166 expression. (A) ChiP analysis of the binding of STAT3 to the CD166 promoter. Data are from 1 representative experiment from a total of 3 that showed very similar results in each. (B) CD166 expression on LSK and LSK48⁻150⁺ cells from WT and STAT3^−/− mice (n = 3 mice per group). Data shown are from 1 mouse/genotype. Identical results were obtained from the other 2 mice. LSK cells were gated from column “A” and LSK48⁻150⁺ were gated from column “B.” (C) CD166 expression on BM-derived Lin⁻ cells treated with STATTIC for 24 hours. Percentage of CD166⁺ cells declined from 53% (solid line plot) to 17% (dotted line plot) after treatment. (D) Expression of CD166 in human MOLT4 cells transduced with scrambled (Src), CD166, or STAT3 short hairpin RNA. Short hairpin RNA was cloned into Age1 and EcoR1 sites of the MISSION vector (Sigma), which was then selected by puromycin.