Page 171 - 2019_12-Haematologica-web

Platelet HIF-2α induces a prothrombotic state
E
H
FG
IJ
AB
CD
Figure 1. Enhanced expression of HIF-2a in human platelets under hypoxia and upon stimulation with physiological agonists. (A, B) Western blot analysis showing expression of HIF-2a in platelets exposed to either hypoxia (1% O2, 5% CO2, and 94% N2) for the indicated times (A) or agonists (thrombin, Thr, 1 U/mL; ADP, 10 μM; collagen, Coll, 10 μg/mL) under non-stirring condition for 10 min at 37°C (B). (C, D) Corresponding densitometric analyses of HIF-2a normalized to β-actin (n≥3). (E, F) Expression of HIF-2a in platelets pretreated or not with puromycin (Puro, 10 mM) and then exposed to hypoxia (E) or thrombin (F). (H, I) Corresponding densito- metric analyses of HIF-2a normalized to β-actin ((n≥4). (G) HIF-2a expression of platelets pretreated with SB202190 (SB, 20 μM and 40 μM). (J) Corresponding den- sitometric analysis of HIF-2a normalized to β-actin (n=6). Data are represented as the mean ± standard error of mean of at least three different experiments. *P<0.05; **P<0.01; #P<0.05; ##P<0.01, analyzed by the Student t test.
haematologica | 2019; 104(12)
2485