Identification of Lhx5 binding sites

Lhx5 mutant mouse embryos show loss of a neuronal nucleus of the brain, called the mammillary body and essential for memory. We found out that this is caused by a loss of appropriate specification of mammillary neurons and we identified in a microarray study downstream targets responsible for the observed phenotype. Here we wanted to find out which of our identified targets are under direct control of Lhx5. Overall design: In order to identify the direct targets of Lhx5 we decided to use a ChIP-Seq approach. For this approach we generated a stably transfected Neuro2a cell line that expresses a FLAG-tagged version of Lhx5 under the control of the Tet-ON system. As a control we transfected Neuro2a cells with a construct that only expresses the FLAG-tag.