Fig. 2

Endoglin⁺ EMPs increase cell proliferation, angiogenesis and TGF-β signalling in HPAECs. a Proliferation and (b) angiogenesis in HPAECs incubated with unmodified (full), VE cadherin- depleted (VE⁻) or endoglin- depleted (endoglin⁻) MP fractions, in the absence or in the presence of TGF-β (10 ng/mL; 18 h), as indicated. Graph in (c) and a corresponding, representative western blot in (d) show phosphorylation changes of smad 1/5/8 in cells pre-incubated with MPs for 1 h and then treated with 10 ng/mL TGF-β for 1 h. *P < 0.05; **P < 0.01; ***P < 0.001, comparisons with healthy controls; ^# P < 0.05, ^## P < 0.01 comparisons, as indicated. N = 3. e Changes in endoglin expression in HPAECs cultured for 18 h with unmodified (full), VE⁻ and endoglin⁻ MPs from healthy volunteers or CTEPH patients. f BMPRII expression in HPAECs incubated with MPs for 18 h. g Western blot showing VE-cadherin (CD144) and endoglin (CD105) levels in MP fractions, as indicated. Briefly, plasma MPs were incubated o/n with Dynabeads coupled with antibodies against CD144 or CD105. The beads with captured proteins were collected and washed in PBS. Then the beads and, full MP fraction and endoglin/VE-cadherin-depleted fractions were resolved by electrophoresis. Changes in endoglin and VE-cadherin levels were studied by western blotting