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

Figure 2. P2Y1, 12, and 13 receptors are not involved in ADP-induced nucleolin down-regulation.

Figure 2

(A, B) The mRNA levels of P2Y1, 12, and 13 receptors in HUVEC (A) and HAEC (B). Cells were treated with the indicated concentrations of ADP for 24 h. The mRNA levels of P2Y1, 12 and 13 receptors were detected by RT-PCR. β-actin transcript was detected as a loading control. (C) The mRNA levels of P2Y1, P2Y12, and P2Y13 receptors in 5–8F cells. (D) The effect of P2Y1 receptor inhibitor MRS2179 (M2179) on ADP-induced down-regulation of nucleolin protein. HUVEC, pre-treated with the indicated concentrations of MRS2179 for 1 h, were re-treated with 100 µM ADP for 72 h. The nucleolin protein levels were detected by western blot. β-actin transcript was detected as a loading control. (E) The effect of P2Y12 receptor inhibitor PSB0739 (PSB) on ADP-induced down-regulation of nucleolin protein. (F) The effect of P2Y13 receptor antagonist MRS2211 (MRS) on ADP-induced down-regulation of nucleolin. (G, H) The effect of MRS2211 on the nucleolin protein levels in HUVEC (G) and HAEC (H). (I, J) The effect of P2Y13 receptor agonist 2-MeSADP on nucleolin protein levels in HUVEC (I) and HAEC (J). (K) 2-MeSADP did not regulate over-expressed nucleolin protein. Nucleolin-over-expressed HUVEC were treated with the indicated concentrations of ADP for 72 h. The GFP-nucleolin expression was detected by fluorescence microscope.