Figure 4.

Effects of ANTXR2 on cell adhesion, migration, and angiogenesis. (A) ANTXR2 was knocked down by siRNA (40 nM) for 48 h in both eutopic and ectopic stromal cells (n = 3). Real-time cell analysis (RTCA) was used to analyze adhesion ability change in those cells. (B) Eutopic stromal cells (n = 4) with single-knockdown of EZH2 (siEZH2) or double-knockdown of EZH2 and ANTXR2 (siEZH2 + siANT) were analyzed by RTCA to determine cell adhesion ability. (C) Eutopic stromal cells pre-treated with control or ANTXR2 siRNA (40 nM) for 24 h and subsequently cultured under normoxia (Nor) or hypoxia (Hypo) for another 24 h. Cells were then subjected to adhesion ability analysis by RTCA (n = 4). (D) Eutopic stromal cells were cultured under normoxia (Nor) or hypoxia (Hypo) with or without 1 μM PGG treatment for 24 h. Cells were then subjected to adhesion ability analysis by RTCA (n = 4). (E) Ectopic stromal cells were plated in the dish coating with 0.5 mg/mL collagen type IV (Col IV) or (F) 0.5 mg/mL Laminin and treated with different doses of PGG (0.2, 1, and 5 μM) for 24 h. Eutopic stromal cells were cultured in un-coated dish as a control. Cells were then subjected to adhesion ability analysis (n = 3). Asterisk (*) indicates P < 0.05. (G-K) ANTXR2 in ectopic stromal cells were knocked down by siRNA for 48 h or treated with different doses of PGG for 48 h and (G, H) cell proliferation (n = 5), (I, J) migration (n = 4), and (K) tube formation (n = 3) abilities were analyzed. Asterisk indicates P < 0.05. Results were presented as means ±SEM. Scale bar: 200 μm.