Polyethylene glycol 400

CAB 171-15 and C-A-P polymers were obtained from Eastman Chemical Company, Kingsport, TN, USA. DFS were purchased from Leap Chem, Hangzhou, China. Acetone, acetonitrile (ACN), monobasic potassium phosphate, croscarmellose sodium (CCS), magnesium stearate (MGS), polyethylene glycol 400 (PEG, Fisher Scientific, Asheville, NC, USA), hydroxypropyl cellulose (HPC, MW 100,000, Sigma-Aldrich, St Louis, MO, USA), lactose monohydrate (LMH, Supertab 145D, Mutchler Inc, Harrington Park, NJ, USA), and microcrystalline cellulose (MCC, Vivapur^® 102, JRS Pharma, Patterson, NY, USA) were used as received. In-house water (18 MΩ.cm, Millipore Milli-Q Gradient A-10 water purification system) was used in the study.

The GSK-3 inhibitor CT99021 was purchased from Cayman and Tocris (Cayman: cat# 13122; Tocris: cat# 4423). The drug was firstly dissolved in 100% DMSO and was then diluted in a mixture of polyethylene glycol 400 (Fisher) and saline solution (0.9% NaCl). The final solution consisted of 10% DMSO, 45% polyethylene glycol 400 and 45% saline. The solution containing CT99021 was administrated by intraperitoneal injection (i.p.) at 25 mg/kg. Drug administration took place every day 1 h prior to the behavior tests apart from delayed match-to-place T-maze (DMTP T-maze). For the DMTP T-maze, drug injection was carried out 30 min prior to the test. The experimenter was blinded to whether the mouse received CT99021 or vehicle. Ketamine (100 mg/ml) was diluted to 1 mg/ml in saline solution and injected (i.p.) at 20 mg/kg.

All animal procedures were approved and carried out in accordance with relevant guidelines and regulations established by the Institutional Animal Care and Use Committee at Vanderbilt University. Briefly, Notch1^+/- and Htr2b^-/- mice were crossed to create Notch1; Htr2b double mutant animals [26 (link), 27 (link)]. In a separate cohort, Alzet minipumps (model 1004) delivering either the high-affinity 5-HT_2B antagonist SB204741 (Tocris Biosciences; 1 mg kg^-1d^-1) or vehicle (50% dimethyl sulfoxide [Sigma-Aldrich] and polyethylene glycol-400 [Fisher Chemical]) were implanted subcutaneously in Notch1^+/- mice at 4 months of age (Fig 1). SB204741 is a well-validated and specific 5-HT_2B receptor antagonist commonly used in pharmacological studies targeting this ligand [28 (link), 29 (link)]. SB204741 dosing varies widely across the literature, but we use 1 mg kg^-1d^-1 to allow for longer time course administration with good results [30 ].
All mice were fed a 1% cholesterol Western diet (TestDiet 5TJT) for a total of 3.5 months beginning at 2.5 months of age. Food and water were provided ad libitum. Mice were sacrificed at 6 months of age using carbon dioxide inhalation followed by cervical dislocation before harvesting biological samples for processing and analysis.

The ferrite feedstock was prepared by mixing a commercial NiCuZn ferrite powder (LSF90, Powder Processing & Technology, LLC, Valparaiso, IN, USA, average particle size 0.8 μm), monomer binder (pentaerythritol tetraacrylate, Sigma-Aldrich Co. LLC, St. Louis, MO, USA), and diluent (polyethylene glycol 400, Alfa Aesar, Haverhill, MA, USA). The weight ratio of ferrite powder to monomer binder was 19. The solid loading (fraction of ferrite powder in the feedstock) was 78 wt %.
The silver feedstock was made by tailoring a commercial silver paste (nanoTach-X, NBE Tech, LLC, Blacksburg, VA, USA). The addition of ceramic powders into metal paste has been shown to be effective in mitigating shrinkage mismatch between ceramic and metal layers during co-firing of multilayer monolithic thick films [22 (link),23 (link),24 (link)]. Thus, to adjust the shrinkage of the silver feedstock, LSF90 ferrite powders were added at different concentrations of 1 wt %, 5 wt %, and 10 wt % into the silver paste. To homogenize the mixture in a container, zirconia milling media of 5 mm diameter and at around 1:1 media-to-content weight ratio was added and then shaken for 5 min in a high-speed vibrating ball miller (MSK-SFM-3, MTI Corp., Richmond, CA, USA).