Figure 2.

Schematic illustration of neuronal nuclei isolation and chromatin immunoprecipitation. (a) Isolation of neuronal nuclei using FACS. Upper panel shows the schematic process of each step, and lower panels show representative data. Brain samples were homogenized and subjected to density gradient centrifugation to obtain crude nuclear isolates. All the nuclei were stained with 7-AAD, and separation of neuronal and non-neuronal nuclei was performed by Alexa488-conjugated anti-NeuN antibody. FACS separation was performed. The lower panels are immunofluorescence images of neuronal and non-neuronal nuclei and neuronal/non-neuronal nuclei isolation using FACS. (b) Schematic diagram of ChIP. The obtained nuclei were sonicated to fragment the genomic DNA into nucleosome units. The nucleosomes with the target histone modifications were captured using an antibody against the target modification. The captured DNA was purified and subjected to library preparation and next generation sequencing.