Sr144528

Androgen-insensitive
(DU145 ACC 261, PC-3 ACC 465) and androgen-sensitive (LNCaP ACC 256)
human prostate cancer cell lines were purchased from the German Collection
of Microorganisms and Cell Cultures (DSMZ). Noncancerous human prostate
epithelial cell lines (PWR-1E CRL-11611, RWPE-1, CRL-11609) were purchased
from the American Type Culture Collection (ATCC). DU145, PC-3, and
LNCaP cells were cultured using RPMI 1640 medium with GlutaMAX (Gibco),
supplemented with 10% fetal bovine serum (FBS) (Gibco). PWR-1E and
RWPE-1 cells were cultured using keratinocyte serum-free medium supplemented
with l-glutamine, bovine pituitary extract (50 mg/L), and
epidermal growth factor (5 μg/L) (Gibco). Cells were maintained
in a humidified 5% CO₂ incubator at 37 °C.
CBD
was provided by GreenLight Pharmaceuticals. CBD purity of >99.7%
was
confirmed by convergence chromatography. SR141716 (CB₁ antagonist),
SR144528 (CB₂ antagonist), capsazepine (TRPV1 antagonist),
LPI (GPR55 agonist), and staurosporine (apoptosis positive control)
were purchased from Sigma-Aldrich. CBD, SR141716, SR144528, and capsazepine
were dissolved by using dimethyl sulfoxide (DMSO) (PanReac AppliChem).
LPI was dissolved using sterile dH₂O. All drug compounds
were stored according to the manufacturer’s instructions.

Lipopolysaccharide (LPS), interferon-γ (IFN-γ), JWH-133 (JWH), and SR-144528 (SR) were purchased from Sigma-Aldrich (St Louis, MO, USA). N-Methyl-D-aspartate (NMDA), MK-801 (MK), and forskolin (FK) were purchased from Tocris (Bristol, UK). Tau and p-Tau proteins were kindly provided by Prof. J. Avila (CBM, UAM-CSIC, Madrid, Spain). Detailed descriptions of the elaboration and processing of proteins can be found elsewhere [42 (link), 43 (link)]

Ultra-pure grade chemicals were used: Progesterone (4 pregnene-3,20-dione, P₄), lipopolysaccharide (LPS, from Escherichia coli 055:B5), CP55,940, and SR144528 (CB2 antagonist) were purchased from Sigma-Aldrich Corp. (St. Louis, MI, USA). ACEA (CB1 agonist) and AM281 (CB1 antagonist) were obtained from Tocris Bioscience (Bristol, United Kingdom), and JWH-015 (CB2 agonist) was obtained from Enzo Life Sciences (Plymouth Meeting, PA, USA). [³H]CP55,940 was obtained from Perkin-Elmer Life Sciences, Inc. (Boston, MA, USA).

Arachidonyl-2′-chloroethylamide hydrate (ACEA; CB1 agonist), Rimonabant hydrochloride (SR141716A; CB1 antagonist/inverse agonist), GW833972A (CB2 agonist), SR144528 (CB2 antagonist/inverse agonist), and dimethyl sulfoxide (DMSO) (Sigma-Aldrich, St. Louis, MO, USA) were used. These cannabinoid receptor agonists and antagonists were dissolved in DMSO at a concentration of 20 mM and stored at −20 °C until used.
Saponin (Amresco, Solon, OH, USA) was used. Carboxyfluorescein succinimidyl ester (CFSE), brefeldin A, and monensin were obtained from Sigma-Aldrich. RPMI 1640 medium and fetal bovine serum (FBS) (Gibco, Grand Island, NY, USA) were used. Ficoll-Hypaque solution (IsoPrep)(Robbins Scientific Corporation, Sunnyvale, CA, USA) was used. Finally, 7-AAD solution was purchased from BioLegend.