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. 2014 Oct 7;9(10):e110101. doi: 10.1371/journal.pone.0110101

Figure 4. Nucleolin protein down-regulation is involved in ADP-induced cell apoptosis.

Figure 4

(A) ADP induced cell detachment in HUVEC. Cells were treated with indicated concentrations of ADP for 72 h. Medium was replaced with fresh medium containing ADP every day. The cells in representative fields were photographed. (B) ADP induced apoptosis in HUVEC. Cells were treated as same as (A). Unfixed cells were stained with FITC-annexin V/PI. Cell apoptosis was measured by flow cytometry analysis. (C) ADP induced time-dependent cell apoptosis. HUVEC were cultured with 100 µM ADP for indicated time periods. Medium was replaced with fresh medium containing ADP every day. Cell apoptosis was measured as same as (B). * P<0.05 compared with the control group. (D) MRS2211 (MRS) did not reverse ADP-induced cell death. HUVEC, pre-treated with the indicated concentrations of MRS2211 for 30 min, were re-treated with 100 µM ADP for 72 h. Cell death was measured by FITC-annexin V/PI staining. * P<0.05 compared with the control group. (E) Nuleolin-over-expression reversed ADP-induced cell death. Nucleolin-over-expressed HUVEC were treated with the indicated concentration of ADP for 72 h. Medium was replaced with fresh medium containing ADP every day. Cell death was measured by FITC-annexin V/PI staining. Mock-transfected HUVEC were used as control. * P<0.05 compared with the control groups.