St1936

Mice received intraperitoneal injection of ST1936 (10 mg/kg, Tocris Bioscience) or SB258585 (5 mg/kg, Tocris Bioscience) four times at E6, E9, E12, E15. Briefly, newborn mice were deskinned, eviscerated, and immersed in 100% ethanol for 24 h. The samples were fixed in acetone for 24 h and then stained for 24 h in a solution containing 0.1% Alizarin Red, 0.3% Alcian Blue, acetic acid, and 70% ethanol (1:1:1:17, v/v/v/v). They were then transferred to a solution of 1% KOH in 20% glycerol until clear and then stored in glycerol.

Fluoxetine, TAPI-1, SB271046, SB742457, L-685,458 and roscovitine were purchased from Selleck Chemicals. Amitriptyline hydrochloride, ST1936 and SB258585 were from Tocris Bioscience. Protriptyline hydrochloride, amoxapine, trimipramine maleate, SB215505, SB206553, cAMP, L-ascorbic acid and DAPI were purchased from Sigma. Escitalopram oxalate was purchased from Lundbeck. Sertraline hydrochloride was from Pfizer. BACE inhibitor IV (BSI IV) was from Calbiochem. Recombinant human BDNF, GNDF, and IGF-I were from Peprotech. CellTiter-Glo was from Promega. Immunoblotting was performed with the following antibodies: anti-ADAM10 (Ab1997, Abcam), anti-BACE1 N-termimus (AP7774b, Abgent), anti-APP-CTF (A8717, Sigma), anti-actin (A2066, Sigma), anti-CDK5 (sc-173, Santa Cruz), and Rabbit anti-β-arrestin-1/2 (A1CT) antibody was a kind gift from Dr. Robert J. Lefkowitz. Immunofluorescence staining was performed with the following primary antibodies: anti-Tuj1 (801201, BioLegend), anti-Sox2 (sc-17320, Santa Cruz), anti-Map2 (Ab5622, Millipore), and anti-GFAP (sc-6170, Santa Cruz).