Ppads

After determining the ADT, either LSKL (50, 100, or 200 μg, Sigma, St. Louis, MO, USA), PPADS (10, 20, or 30 μg, Abcam, Cambridge, MA, USA), Reactive Blue 2 (20 μg, Bomei, China), or saline in 5 μl volume was injected once daily into the right lateral cerebral ventricle, over a period of 10 min, using a disposable dental needle. The needle was held in place for 5 min before being slowly retracted. siRNA was designed and synthesized by Tuoran Biological Technology Co., Ltd. (Shanghai, China) using the following oligonucleotide sequences: 5’-GCCAGUAUGUUUACAACGUdTdT-3’ and 5’-ACGUUGUAAACAUACUGGCdTdT-3’. Negative controls were produced using the following oligonucleotides: 5’-UUCUCCGAACGUGUCACGUTT-3’ and 5’- ACGUGACACGUUCGGAGAATT-3’.
Increasing doses of siRNA (0.5, 1.5, 2.5 μg) or negative control (control; all 5 μl) were injected into the right lateral cerebral ventricle every other day. The injection was performed over a period of 10 min, after which the needle was held in place for 5 min. Behavioral seizure activity and EEGs in the amygdala were recorded after kindling stimulation every day. The ADT was then determined again after the stimulations.