Fig. 6. Math1 bound the Mbh1 enhancer containing the E-box in the spinal cord. (A) ChIP demonstrating binding of endogenous Math1 to the enhancer. Formaline-cross-linked chromatin from the E10.5 spinal cord was incubated without (–) and with rabbit polyclonal antibodies against Math1 and neurofilament. (B) Misexpressed Math1 also bound the enhancer. One day after electroporation of Math1-HA, cross-linked chromatin from the E12.5 spinal cord was incubated without (–) and with a rat monoclonal anti-HA antibody and IgG. Immunoprecipitates were analyzed by PCR using primers specific to the Mbh1 enhancer (upper panels) and to the region 5.8 kb upstream of the E-box as a negative control (lower panels). Each input represents DNA purified from the chromatin before immunoprecipitation. These data are representative of two (A) and four (B) independent experiments. Arrows and arrowheads indicate the amplified 502 bp and 546 bp DNA fragments, respectively.