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. 2020 May 14;8(1):e000610. doi: 10.1136/jitc-2020-000610

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© Author(s) (or their employer(s)) 2020. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See http://creativecommons.org/licenses/by-nc/4.0/.

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Production of adenosine during coculture of CD39⁺ JJN-3 HMCL and CD73⁺ HS-5. (A) JJN-3 and HS-5 cells, cultured separately or together, were incubated as described in online supplementary methods with 100 µM ATP for indicated time periods. Adenosine concentration in supernatants was measured by HPLC-MS. Medium (CM) from JJN-3 and HS-5 cells cultured in the absence of ATP was used as control. (B) POM-1 and anti-CD73 inhibited adenosine generated by JJN-3 and HS-5. HS-5 cells were incubated with 100 µM ATP, JJN-3, or POM-1-pretreated JJN-3, 150 µg/mL anti-CD73, or isotype for 1 hour. Then the supernatants were collected and adenosine concentration was measured by HPLC-MS. Data were means±SD. Statistical differences were calculated using Student’s t-test. Three independent experiments were performed.