Pi3k specific inhibitor ly294002

Intracerebroventricular drug administration was performed as previously described [21] (link). Briefly, rats were placed in a stereotaxic apparatus under anesthesia with isoflurane (4% induction, 2.5% maintenance). The needle of a 10-μl Hamilton syringe (Microliter701; Hamilton Company, Reno, NV, USA) was inserted into the right lateral ventricle through a burr hole using the following coordinates relative to bregma: 1.5 mm posterior, 1.0 mm lateral, and 3.2 mm below the horizontal plane of the bregma. For LRP1 in vivo knockdown, a total of 500 pmol LRP1 siRNA duplexes (Thermo Fisher Scientific, Waltham, MA) were dissolved in 5 μl RNase free suspension buffer and then infused into the right lateral ventricle via a pump with the rate 1 μl/min at 48 h before SAH induction. The same volume of scrambled (Scr) siRNA (Thermo Fisher Scientific, Waltham, MA) was used as a negative control. PI3K-specific inhibitor LY294002 (Selleck Chemicals, Houston, USA) was prepared at 50 mmol/L in PBS (contains 25% DMSO) with a total volume 5 μl. The LY294002 was injected 30 min before SAH. The same volume of 25% DMSO was used as a negative control for LY294002. After injection, the needle was kept in place for an additional 5 min and retracted slowly. Then, the burr hole was sealed with bone wax immediately, and the rats were allowed to recover after sutures.