Lmx1a works through a well-conserved binding site in the DAT promoter région. a. ChIP-Real time PCR analysis on DAT promoter region. In vitro differentiated cells were transduced with HA-tagged Lmx1a-expressing retrovirus and fixed for ChIP at ND3. ChIP fragments were immunoprecipitated with normal rabbit IgG or anti-HA antibody and analyzed by qPCR. The average of three independent ChIP analyses (n=3, mean±SEM, p<0.05) are presented. b. E13.5 VMs were dissected and used for ChIP using Lmx1a antibody. Binding of Lmx1a to the DAT promoter was assayed by qPCR (n=3 from independet ChIP reactions, mean±SEM, p<0.05). c. Reporter construct containing multiple copies of DAT-A mediates transactivation by Lmx1a (n=3 from independent transfection reactions, mean±SEM, p<0.05). Reporter constructs were cotransfected with either empty vector or Lmx1a expression vector to human neuroblastoma SK-N-BE(2)C cells and Luciferase activaty was measured 24–48 hours after transfection and normalized by β-galactosidase activity.