Fig 8. Inhibition of ATR activation inhibits the NS1-induced G2-phase arrest of CD36⁺ EPCs.

(A) Cell cycle analysis. CD36⁺ EPCs were treated with VE821 at 3 h prior to NS1-expressing lentivirus transduction or mock transduction. After 48 h, cells were collected, and the cell cycle phase of NS1-expressing cells (selected by staining with anti-Flag) was examined by flow cytometry. (B) Statistical analyses. The percentage of cells treated with DMSO or VE821 that were at G1-, S-, and G2-phase is depicted in color. The numbers shown are the percentages of the cells at G2-phase, and are statistically compared as indicated. ** P<0.01. (C-E) Western blot analysis. (C) CD36⁺ EPCs were either treated with DMSO or VE821 at 3 h prior to lentivirus or mock transduction. After 48 h, cells were collected for Western blotting to detect ATR(pT1989). (D&E) CD36⁺ EPCs were transduced with Lenti-NS1 or Lenti-NS1^mTAD2. After 48 h, cells were collected for Western blotting to detect ATR(pT1989), CHK1(pS345), CDC25C(pS216), CDK1(pY15), and β-actin. Cells treated with HU or Noco at 24 h prior to analysis were used as controls.