Splenocytes from wild type (WT) mice and P2RY12-/- mice were isolated and cultured in 96-well plates at a density of 1×106 cells per well, then stimulated with ConA (2 μg/ml) (Sigma-Aldrich) for 12 hr or 24 hr and co-cultured with the P2RY12 inhibitors clopidogrel (1, 3, 10 μM) or ticagrelor (1, 3, 10 μM), HK2 inhibitor 3-bromopyruvate (3-BrPA, MedKoo Biosciences Inc., Morrisville, NC, USA), proteasome inhibitor MG-132 (Med-Chem-Express LLC, Monmouth Junction, NJ, USA), lysosomal inhibitor bafilomycin A1 (BafA-1) (Med Chem Express LLC) or Akt activator SC79 (Med-Chem-Express LLC). The cells were analyzed by flow cytometry and supernatants were collected for measurement of cytokines.
3 bromopyruvate 3 brpa
Manufactured by MedKoo Biosciences
Sourced in United States
3-bromopyruvate (3-BrPA) is a chemical compound used in research applications. It functions as a glycolysis inhibitor.
Automatically generated - may contain errors
2 protocols using 3 bromopyruvate 3 brpa
1
Splenocyte Activation Assay with P2RY12 Inhibitors
2
DDP-induced Cytotoxicity in LUAD Cells
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The primary LUAD, A549 and A549/DDP cells were plated in a 96-well plate (3×103 cells/well) and cultured overnight at 37°C. The following day, primary LUAD cells were exposed to 10 µM DDP (Selleck Chemicals) for 48 h. A549 cells were transduced with the PRDM14 overexpression vector (oePRDM14) and treated with 10 or 20 µM DDP or 20 µM antiglycolytic agent 3-Bromopyruvate (3-BrPA; MedKoo Biosciences, Inc.) for 48 h at 37°C. A549/DDP cells were transfected with shPRDM14 and treated with 10 or 20 µM DDP for 48 h at 37°C. Following treatment, 10 µl CCK-8 solution (Signalway Antibody LLC) was added to each well for 1 h according to the manufacturer's instructions. The optical density (OD) value of each well was read on a microplate reader at 450 nm to calculate relative cell viability inhibition rate as follows: Viability inhibition rate = (ODvehicle + vector - ODDDP + vector)/ODvehicle + vector ×100%.
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