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Pumps delivered their contents into the right lateral cerebral ventricle via a brain infusion kit (ALZET).
A sterile brain infusion cannula (28-gauge; Brain infusion kit 1; Alzet, Palo Alto, CA) was stereotaxically implanted into the left third ventricle.
Osmotic pumps (Alzet, model 2004) with a brain infusion kit (Alzet) were filled with reagents or sterile saline and placed into sterile saline at 37°C for at least 40 hours, according to the manufacturer's instructions.
The infusion kit was implanted into the right cerebral ventricle at coordinates relative to bregma: AP −1.0 mm, L +1.5 mm and V 4 mm below the dura and fixed with dental cement at the skull.
Osmotic pumps were filled with either ACSF or PPADS, conditioned for 14 hours at 37°C in sterile saline solution, and afterwards connected with a rat brain infusion kit (Alzet®, Charles River).
The Alzet Brain Infusion Kit 3 (Alzet Corp., CA) and the Alzet osmotic mini-pump (Alzet 1002, Alzet Corp., CA), were used to deliver rhGas6 or PBS (vehicle) directly into the corpus callosum.
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For intrathecal infusion, purified ephrinB3 extracellular domain proteins (ephrinB3) at 100 μg/ml or vehicle 1 × phosphate buffer saline (PBS, pH7.4, Gibco, Langley, OK, USA) was infused in the injury epicenter using a micro-osmotic pump (Alzet, Durect Corp ,Cupertino, CA, USA; model 1007D) assembled according to the manufacturer's recommendations and connected to Brain Infusion kits #3 (Alzet).
IPS includes electroconvulsive therapy (ECT), Ketamine Infusion Therapy (KIT) and Transcranial Magnetic Stimulation (TMS).
The amplified 2.83-kbp fragment of CE0634-CE0635 was fused to BglI-digested pCH using the InFusion Cloning Kit (Clontech Laboratories Inc ,Mountain View, CA, USA).
The SKA2 promoter luciferase construct was generated using the Infusion Cloning kit.
PAI-1 3′-UTRs were PCR amplified from BAC clone RP11-213E22 and inserted into the pMIR vector using the Infusion cloning kit (Clontech) and primers listed in Table 1.
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