Primary mouse hepatocytes cultured in six-well plates were washed with HPSB buffer containing 0.2% FFA-free BSA, 2 mM glucose, and 2 mM pyruvate and then incubated in the same buffer for 3 hours in a 5% CO2 incubator. Supernatant and cell pellets were collected and saved for further assays. Extracellular adenosine levels were determined in the hepatocyte-conditioned media using a colorimetric assay kit (BioVision, catalog no. K327). Plasma membrane fraction of the cell pellets (Abcam, catalog no. ab65400) was subjected to ADA activity assays using a colorimetric assay kit (BioVision, catalog no. K321).

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