Adenosine 5 triphosphate disodium salt hydrate

From Sigma-Aldrich (St. Louis, MO, USA): Adenosine 5′-triphosphate disodium salt hydrate (ATP, A1852), DMEM (D6429), Hanks′ Balanced Salt Solution (HBSS, H4641), HEPES sodium salt (H7006), Monensin (M5273), Nystatin (N6261), Amiloride (PHR1839), Xestospongin C (X2628), MRS-2179 (M3808), U73122 (U6756), Thapsigargin (T9033), Ryanodine (559276), Carbenoxolone (C4790), ¹⁰Panx1 (SML2152), Bafilomycin A1 (19-148), Apyrase (A6237), Gap26 (SML3074). From Molecular probes (Eugene, OR, USA): Hoechst 33,342 (H1399), Propidium iodide (P1304MP). From Evrogene (Moscow, Russia): MMLV reverse transcriptase (SK022S), SYBR Green I PCR Master Mix (PK147L); Triethylammonium salt (TNP-ATP, Ann Arbor, MI, USA, 20902). From Thermo Fisher Scientific (Waltham, MA, USA): Fura-2AM (Cat. #F1221), Fetal Bovine Serum (10099141). Selenium nanoparticles (kindly provided by Dr. S.V. Gudkov, Prokhorov General Physics Institute of the Russian Academy of Sciences, Moscow, Russia).

AsPC-1 cells, a cell line derived from pancreatic ductal adenocarcinoma [30 (link)] and PS-1 cells, which are immortalised human pancreatic stellate cells [31 (link)] were a kind gift from Prof. Hemant Kocher (Barts Cancer Institute, London, UK). Cells were cultured using RPMI-1640 medium (Gibco, Amarillo, TX, USA) for AsPC-1 cells and DMEM-12 (Sigma-Aldrich, St. Louis, MO, USA) for PS-1 cells. Media was supplemented with 10% fetal bovine serum (Sigma-Aldrich, St. Louis, MO, USA) and cells were kept at 37 °C and 5% CO₂ in a humid incubator. To study the effect of agonist and antagonist treatment, AsPC-1 cells were treated with 100 µM of the P2Y₂ agonist, adenosine 5′-triphosphate disodium salt hydrate (Sigma-Aldrich, St. Louis, MO, USA) and 5 µM of the P2Y₂ antagonist AR-C 118925XX (Tocris, Bristol, UK) for 1 h before fixation.

The HTLV-1-infected cell lines MT1, MT2, and MT4 were purchased from JCRB Cell Bank. The HTLV-1-infected cell line MJ, and the T-cell lymphoma cell lines Jurkat and EG7-OVA were provided by ATCC (VA, USA). All cell lines are identified based on short tandem repeat profiles by providers, and mycoplasma contaminations were denied both by providers and at our laboratories. Cell lines and PBMCs were cultivated using RPMI 1640 Medium (Nacalai Tesque, Kyoto, Japan) supplemented with 10% fetal bovine serum (FBS), and Penicillin–Streptomycin Mixed Solution (×100) (Nacalai Tesque). Mouse anti-CD4, CD8, CD7, CD39, CD73, and CD26 antibodies were purchased from Biolegend (San Diego, CA, USA), and anti-CADM1 antibody was purchased from MBL Inc. (Woburn, MA, USA). adenosine 5′-triphosphate disodium salt hydrate, adenosine 5′-monophosphate hydrate, and adenosine were purchased from Sigma Aldrich (now Merck KGaA, Darmstadt, Germany). Polyinosinic-polycytidylic acd sodium salt (Poly(I:C)) was purchased from R&D systems (Minneapolis, MN, USA). The CellTrace^TM Violet Cell Proliferation Kit was purchased from Thermo Fisher Scientific (Waltham, MA, USA).

ATP was purchased in the form of adenosine 5-triphosphate disodium salt hydrate (Sigma A7699).

Adenosine 5′-triphosphate disodium salt hydrate (≥99%), chlorophenol red-β-d-galactopyranoside (CPRG), β-nicotinamide adenine dinucleotide phosphate hydrate (NADP⁺), magnesium chloride hexahydrate (≥99%), d-(+)-glucose (≥99%), dimethyl sulfoxide (ACS reagent (≥99.9%)), d-glucose-6-phosphate sodium salt, N-(2-hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid) (HEPES, ≥99.5%), imidazole (96.0%), Terrific broth modified, and triethanolamine were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM) was purchased from CellGro Technologies, LLC (Lincoln, NE, USA). Leuconostoc mesenteroides glucose-6-phosphate dehydrogenase (LmG6PDH) was purchased from Worthington Biochemical Corporation (Lakewood, NJ, USA). NADPH tetrasodium salt (≥95%) was purchased from Enzo Life Sciences, Inc. (Farmingdale, NY, USA). For the 3-nitro-2-phenyl-2H-chromene analogues, compounds 1–7 were purchased from TimTec, LLC (Tampa, FL, USA), while compounds 8–13 were purchased from ChemDiv, Inc. (San Diego, CA, USA). Ethylenediaminetetraacetic acid (EDTA) tetrasodium salt hydrate (98%), Luria-Bertani (LB) broth, lysozyme (type VI), protease inhibitor tablets (EDTA-free), glycerol, and all other chemicals were purchased from Fisher Scientific (Hampton, NH, USA).

Spermine tetrahydrochloride, silver nitrate (AgNO₃), sodium borohydride (NaBH₄), adenosine 5′-triphosphate disodium salt hydrate (ATP), protein kinase A (PKA, from bovine heart), and dithiothreitol (DTT) were purchased from Sigma–Aldrich (Shanghai, China). Dihydrochloride hydrate (H-89) was obtained from J&K Scientific Ltd. (Shanghai, China). Protein kinase B (PKB) was purchased from Bio-Techne China Co., Ltd. (Shanghai, China). Forskolin, 3-isobutyl-1-methylxanthine (IBMX), and Tris-HCl buffer solution (pH 7.5) were purchased from Sangon Biotech (Shanghai, China). Substrate peptide for PKA (TAMRA-LRRASLG, 98%), and phosphorylated substrate peptide were purchased from GL Biochem (Shanghai, China). Magnesium chloride (MgCl₂), sodium chloride (NaCl), ethylenediaminetetraacetic acid (EDTA), glycerol, and dimethyl sulfoxide (DMSO) were obtained from Shanghai Chemicals Ltd. (Shanghai, China). The human cervical carcinoma cells (HeLa) were purchased from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). All other reagents were of A.R. grade and used as received without further purification. Ultrapure water with a resistivity of 18.2 MΩ·cm was used.