Fig. 3

Expression and ectonucleotidase activity of CD39, CD73 and A2AR. a Cells stained with respective antibodies and MFI values determined by flow cytometry. BM-MSCs n = 3, LA-MSCs n = 7, CB-MSCs n = 5, HUVECs, n = 6 each biological replicates; HeLa n = 3 technical replicates; Plt unst, Plt stim each n = 4 biological replicates. *p < 0.05, **p < 0.01, ***p < 0.001. b Adenosine concentration (circles, left y axis) measured by mass spectrometry in supernatants of cell and platelet cocultures, either unstimulated or TRAP-6 stimulated. In parallel, inhibitory effect on TRAP-6-induced CD62p expression (crosses, right y axis) calculated relative to the TRAP-6 control. BM-MSCs n = 3, LA-MSCs n = 2, CB-MSCs n = 1, HUVECs n = 3 each biological replicates; HeLa n = 1. ***p < 0.001, ****p < 0.0001. c, d Released phosphate (c) or adenosine (d) indicative of ectonucleotidase activity determined after incubating cells with ATP, ADP and AMP in presence of respective inhibitors (CD39, POM-1; CD73, AMP-CP; A2AR, SCH 58261). BM-MSCs, LA-MSCs, CB-MSCs, HUVECs, Plt unst, Plt stim each n = 3 biological replicates; HeLa n = 1. *p < 0.05, ****p < 0.0001. Note different y axis scale for AMP. ADP adenosine diphosphate, AMP adenosine monophosphate, ATP adenosine triphosphate, BM bone marrow, CB cord blood, HUVEC human umbilical vein endothelial cell, LA lipoaspirate, n/a not analyzed, Plt stim stimulated platelets, Plt unst unstimulated platelets, w/o without