RPMI 1640, with L-Glutamine and 25mM HEPES

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	RPMI-1640 (BE12-167F; Lonza) was supplemented with fetal bovine serum (10270106; Thermo Fisher Scientific), AB-serum (H4522; Sigma-Aldrich) where indicated, 100 U/ml penicillin, 100 mg/ml streptomycin (DE17-603E; Lonza), 2 mmol/l l-glutamine (BE17-605E; Lonza), 25 mmol/l HEPES and 1 mmol/l sodium pyruvate

Protocol tips
RPMI-1640 (BE12-167F; Lonza) was supplemented with fetal bovine serum (10270106; Thermo Fisher Scientific), AB-serum (H4522; Sigma-Aldrich) where indicated, 100 U/ml penicillin, 100 mg/ml streptomycin (DE17-603E; Lonza), 2 mmol/l l-glutamine (BE17-605E; Lonza), 25 mmol/l HEPES and 1 mmol/l sodium pyruvate

Publication protocol

DU145 prostate cancer cells were obtained from the American Type Culture Collection and maintained in culture with regular passaging for less than 6 months. Authentication was carried out by the supplier using cytogenetic, isoenzymatic and DNA profile analysis. The HLA type of DU145 cells is HLA-A03/A33/B50/B57 (Welsh Blood Transfusion Service, Cardiff, UK). The cells were mycoplasma free and tested regularly using a MycoAlert Mycoplasma Detection Kit (LT07-418; Lonza). Irradiation was carried out using a 137Cs-source (with regular dosimetry quality assurance) at a rate of 0.627 Gy/min.

RPMI-1640 (BE12-167F; Lonza) was supplemented with fetal bovine serum (10270106; Thermo Fisher Scientific), AB-serum (H4522; Sigma-Aldrich) where indicated, 100 U/ml penicillin, 100 mg/ml streptomycin (DE17-603E; Lonza), 2 mmol/l l-glutamine (BE17-605E; Lonza), 25 mmol/l HEPES and 1 mmol/l sodium pyruvate (Sigma-Aldrich). FBS was depleted of bovine exosomes by pelleting at 100,000 g for 2 h. Lipopolysaccharide (LPS, L4391), ATP (A6419) and AMP-CP (CD73 inhibitor, M8386) were obtained from Sigma. PGE2, (2292), NECA (1691) and SCH58261 (adenosine A2A receptor inhibitor, 2270) were obtained from Tocris. PGE2-receptor inhibitors AH-6809 (EP2 inhibitor; CAY14050) and AH-23848 (EP4 inhibitor; CAY-19023) were obtained from Cayman Chemicals.

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Manufacturer protocol

Download the product protocol from Lonza for RPMI 1640, with L-Glutamine and 25mM HEPES below.

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