Figure 4.
Figure 4. Gas6-Mer axis is involved in the expression of TM and PAI-1 in ECs. (A,D) The downregulation of TM and upregulation of PAI-1 were induced by exogenous Gas6. The ECs were incubated for 24 hours with exogenous Gas6 (0, 100, 200, 400 ng/mL), followed by western blotting. Representative data are from 4 independent experiments. *P < .05. (B,E) The downregulation of TM and upregulation of PAI-1 were increased by exogenous Gas6, and 5 μmol/L UNC2250, a selective Mer tyrosine kinase inhibitor, significantly inhibited the downregulation of TM and the upregulation of PAI-1 in the ECs, which were then incubated for 24 hours with exogenous Gas6 (100 ng/mL) with or without UNC2250 (5 μmol/L), followed by western blotting. Results are shown as mean ± SD of statistical analyses from 4 separate experiments. *P < .05. (C,F) The downregulation of TM and upregulation of PAI-1 were induced by the exposure of sera isolated from patients with grade III aGVHD at 50% (vol/vol) of the total media, and UNC2250 significantly inhibited the downregulation of TM and upregulation of PAI-1 in ECs. The ECs were incubated for 24 hours with the sera isolated from the patients with or without grade III aGVHD at a level of 50% (vol/vol) of the total medium with or without UNC2250 (5 μmol/L), followed by western blotting. Representative immunoblots from 4 similar experiments are shown. Bars are the means ± SD of quantitative densitometric analyses. *P < .05. (Ga-b) mRNA expressed Mer and Gas6 in human platelets of patients with or without aGVHD. To examine whether the platelet mRNA expression of Gas6 and Mer in human platelets is inhibited by UNC2250, real-time PCR was performed. UNC2250 (5 μmol/L) pretreatment (4 hours) decreased the mRNA expression of Gas6 in human platelets of patients without aGVHD, and did not affect the mRNA expression of Mer. Representative data from 3 experiments are shown. (H-K) The effect of UNC2250 on increased platelet aggregation induced by collagen (2.5 μg/mL) and ADP (5 μmol/L) with or without Gas6 (200 ng/mL). Representative data from 3 similar experiments are shown.

Gas6-Mer axis is involved in the expression of TM and PAI-1 in ECs. (A,D) The downregulation of TM and upregulation of PAI-1 were induced by exogenous Gas6. The ECs were incubated for 24 hours with exogenous Gas6 (0, 100, 200, 400 ng/mL), followed by western blotting. Representative data are from 4 independent experiments. *P < .05. (B,E) The downregulation of TM and upregulation of PAI-1 were increased by exogenous Gas6, and 5 μmol/L UNC2250, a selective Mer tyrosine kinase inhibitor, significantly inhibited the downregulation of TM and the upregulation of PAI-1 in the ECs, which were then incubated for 24 hours with exogenous Gas6 (100 ng/mL) with or without UNC2250 (5 μmol/L), followed by western blotting. Results are shown as mean ± SD of statistical analyses from 4 separate experiments. *P < .05. (C,F) The downregulation of TM and upregulation of PAI-1 were induced by the exposure of sera isolated from patients with grade III aGVHD at 50% (vol/vol) of the total media, and UNC2250 significantly inhibited the downregulation of TM and upregulation of PAI-1 in ECs. The ECs were incubated for 24 hours with the sera isolated from the patients with or without grade III aGVHD at a level of 50% (vol/vol) of the total medium with or without UNC2250 (5 μmol/L), followed by western blotting. Representative immunoblots from 4 similar experiments are shown. Bars are the means ± SD of quantitative densitometric analyses. *P < .05. (Ga-b) mRNA expressed Mer and Gas6 in human platelets of patients with or without aGVHD. To examine whether the platelet mRNA expression of Gas6 and Mer in human platelets is inhibited by UNC2250, real-time PCR was performed. UNC2250 (5 μmol/L) pretreatment (4 hours) decreased the mRNA expression of Gas6 in human platelets of patients without aGVHD, and did not affect the mRNA expression of Mer. Representative data from 3 experiments are shown. (H-K) The effect of UNC2250 on increased platelet aggregation induced by collagen (2.5 μg/mL) and ADP (5 μmol/L) with or without Gas6 (200 ng/mL). Representative data from 3 similar experiments are shown.

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