Genome-wide occupancy of SOX10 in Schwann cells of rat trigeminal neuralgia
Genome-wide occupancy of SOX10 in Schwann cells of rat trigeminal neuralgia
Schwann tissues at REZ of different TN groups were harvested. Schwann cells are washed by PBS twice, and resuspended by 1 mL RIPA buffer, and put on ice for 30 min, followed by highest speed centrifugation for 15 min. The supernant containing with nucleus are transfered to another new tube. whole cell lysates are sonicated to break up the genomic DNA into 200-500 bp fragments. Then, 10% lysates were saved as input and the remaining were incubated with 1 μg IP grade antibody of SOX10 at 4oC overnight. This is followed by incubating for 2 h with Protein A beads (Thermo Fisher Scientific) at 37 °C to pull down the bound DNA fragments. DNA fragments from the pull-down is isolated by ethanol precipitation.For high throughput sequencing, we added 3'-dA overhang to the SOX10 enriched or input DNA and ligated them to the adapter to build a DNA library.