Mouse E12.5 spinal cords were dissected, dissociated, and processed for ChIP assays. The samples were pre-cleared with protein G beads and immunoprecipitated using NFIA antibody (abcam), Sox10 (rabbit-polyclonal), or control IgG (Santa Cruz). The DNA was purified and PCR was preformed using region specific primers. See supplemental information for primer sequences. HEK293 or HEK293T cell lines cells were transfected with pGL3-reporter constructs and a CMV-β-galactosidase vector using Superfect transfection reagent (Qiagen). Cells were harvested and analyzed for luciferase activity; β-galactosidase was used to normalize for transfection efficiency.
Co-immunoprecipitation was performed by transfecting P19 or Oli-Neu cells with Flag-NFIA and/or HA-Sox10; harvested cell lysates were subject to immunoprecipitation using a specific antibody or IgG control and protein G agarose beads. Seesupplemental information for additional information.
Co-immunoprecipitation was performed by transfecting P19 or Oli-Neu cells with Flag-NFIA and/or HA-Sox10; harvested cell lysates were subject to immunoprecipitation using a specific antibody or IgG control and protein G agarose beads. See