Gtx1 15

Human prolactin-releasing peptide (hPRP), ProTx-I, ProTx-II, and GsMTx-4 were purchased from Peptide Institute, Inc. (Osaka, Japan). GTx1-15 was obtained from Alomone Labs (Jerusalem, Israel). Pepsin, elastase, glycine, HCl, and Tris-HCl were purchased from Wako (Osaka, Japan). Trypsin and α-chymoTrypsin were purchased from Sigma-Aldrich (St. Louis, MO, USA) and Tokyo Kasei (Tokyo, Japan), respectively. All enzymes and peptides were dissolved in distilled water to make 100 ng/μL solutions. glycine was dissolved to make 1 M solution and adjusted to pH2.0 with 5 M HCl and 1 M Tris-HCl (pH8.0) was diluted to 500 mM.

The drugs used in the experiments are in below table.
Chemicals and drugs were purchased from Sigma (St. Louis, MO, USA), except GTX1–15 which was purchased from Alomone Labs (Jerusalem, Israel). Bicu, CsCl and8-Br-cAMP were prepared in distilled water, while the other chemicals were dissolved in dimethylsulfoxide (DMSO; Sigma), and diluted to the required concentration in ACSF immediately prior to use, resulting in a maximal DMSO concentration of <0.1%. Application of drugs was achieved by switching a multi-valve, single-output gravity perfusion system made of square glass capillary (0.4 mm in width, Cat.#64–0121, Warner, USA) to the experimental chamber at a speed about 2 ml/min.

GTx1-15 was obtained from Alomone Labs (Jerusalem, Israel). Human plasma was purchased from KOJIN BIO (Saitama, Japan). The protein thermal shift starter kit was from Thermo Fisher Scientific (Tokyo, Japan). RPMI-1640 with L-glutamine and phenol red, sodium hydrogen carbonate, and Tris-HCl were purchased from Wako Chemicals (Osaka, Japan). A human monocytic leukemia cell line, THP-1 (No. JCRB0112), was from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan). Penicillin-streptomycin solution was obtained from Life Technologies Japan Ltd. (Tokyo, Japan). HyClone fetal bovine serum was purchased from GE Healthcare Japan (Tokyo, Japan). Premix WST-1 reagent was purchased from Takara Bio (Shiga, Japan).