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. Author manuscript; available in PMC: 2017 Nov 1.
Published in final edited form as: J Thromb Haemost. 2016 Oct 5;14(11):2261–2273. doi: 10.1111/jth.13477

Figure 7.

Figure 7

Platelet releasates regulate expression, localization and stability of cyclin D1 in EA.hy926 cells. (A) Dose response of platelet releasates (30 min) for inducing the phosphorylation of GSK-3α/β in EA.hy926 cells. PolyP-70 is shown as a control in the last lane. (B) Overexpression of cyclin D1 by boiled (30 min) platelet releasates (plt-rel, 0.1 ratio) is inhibited by co-incubation of platelet releasates with EcPPXc (250 µg/ml) and alkaline phosphatase (ALP 2 units/mL). PolyP-70 is shown as a control in the lanes 2–4. (C) Analysis of cyclin D1 overexpression by boiled (30 min) platelet releasates by the inhibitors of mTOR upstream signaling pathway. With all inhibitors, cells were treated with boiled platelet releasates co-incubated with each inhibitor for 8h. (D) Platelet releasates-mediated nuclear localization of cyclin D1 was measured in EA.hy926 cells co-incubated with ERK1/2 inhibitor (PD-98059), mTORC1 inhibitor (rapamycin), IKKα and IKKβ inhibitors (BAY11-7082 and BMS-345541 respectively). (E) The same as D except that platelet releasates-mediated inhibition of cyclin D1 phosphorylation was monitored. The scale for the microscopic figure is 20µm. The results are shown as mean ± standard deviation of 3 different experiments. *P<0.05; **P<0.01.