Fig. 4
Selective inhibition of NTPDase3 attenuates the extracellular catabolism of A ATP and B UDP in BM-MSCs from Pm women allowed growing for 21 days in an osteogenic-inducing medium. ATP (100 μM) or UDP (100 μM) were added to the culture medium at time zero either in the absence (left-hand-side panels) or in the presence of PSB 06126 (3 µM; middle panels) or hN3-B3S (0.5 µg/mL; right-hand-side panels). 75-μl samples were collected at the indicated times for HPLC analysis to quantify the substrates ATP or UDP (white bars) and their metabolites, namely ADP (black), AMP (grey), adenosine (ADO, red), inosine (INO, orange), hypoxanthine (HX, green) or UMP (grey) and uridine (red). The calculated half-life time (t1/2) for each initial substrate is shown for comparison; aP < 0.05 and bP < 0.01 (two-tailed unpaired t-test) represent significant differences. Right-hand-side box and whiskers graphs represent the effect of PSB 06126 (3 µM) and hN3-B3S (0.5 µg/mL), assessed by comparing the slope variation of ATP or UDP disappearance in the presence of NTPDase3 inhibitors or under control conditions (with no added drugs); zero represents identity between obtained values in the absence and the presence of the inhibitors. Box and whiskers represent pooled data from eight to twelve Pm women (69 ± 3 years old); cells from each individual were tested in triplicate. bP < 0.01 and cP < 0.001 (one-way ANOVA with Sidák’s multiple comparison test, single pooled variance) represent significant differences