Nigericin solution

The operation of intracerebroventricular injection was as previously described (13 (link)). A hole, lateral to the bregma 1.0 mm, was drilled on the skull without puncturing the dura. A needle of a 10-μL Hamilton syringe (Microliter 701; Hamilton Company, Reno, NV, USA) was then stereotactically and vertically inserted into the hole, 3.0 mm under the horizontal plane of the bregma, entering the left lateral ventricle. A total 2 μL volume of recombinant NEK7 protein solution (Sino Biological, Beijing, China, 4 μg dissolved in 2 μL sterile saline), nigericin solution (Sigma-Aldrich, Shanghai, China, 4 μg in 2 μL vehicle, vehicle: 2 μL mixture of 9:1 NS and DMSO), or MCC950 solution (Sigma-Aldrich, Shanghai, China, 2 μg in 2 μL vehicle, vehicle: 2 μL mixture of 9:1 NS and DMSO) was infused at a rate of 0.2 μL/min, 1 h after SAH induction, whereas 500 pmol/2 μL of NEK7 siRNA (Santa Cruz Biotechnology, Santa Cruz, CA, USA, 500 pmol in a 2-μL mixture of 1:1 RNase-free water and lipofectamine) or scrambled siRNA was infused at the identical rate 48 h before SAH induction. The syringe was slowly withdrawn at 10 min after infusion.