Figure 2

EGCG inhibits migration and capillary tube formation by HUVEC cells. (A) Migration of HUVEC cells was assessed using Transwell Boyden chamber containing a polycarbonated filter. HUVECs (4 × 10⁴ cells) were pretreated with AKT inhibitor IV (1 μM) and/or MEK1/2 inhibitor PD98059 (10 μM) for 2 h, followed by treatment with EGCG (40 μM) or DMSO (control). Migration through the membrane was determined after 24 h of incubation at 37°C. Cells that had migrated to the lower chamber were fixed with 90% methanol, stained with giemsa, quantified by counting the number of cells under a microscope. Data represent mean ± SD. * = significantly different from control, P < 0.05. (B), HUVEC cells were treated as described in A. Cells that had migrated to the lower chamber were fixed with 90% methanol, and photographed with a digital camera attached to a microscope. (C), HUVECs (10 × 10⁴) were seeded in 24-well plates containing matrigel, and pretreated with AKT inhibitor IV (1 μM) and/or MEK1/2 inhibitor PD98059 (10 μM) for 2 h, followed by treatment with EGCG (40 μM) or DMSO (control) for 24 h. Capillary tube structures were photographed with a digital camera attached to a microscope. (D), HUVECs cells were seeded and treated as described in C. Capillary tubes were counted under a microscope. Data represent mean ± SD. * = significantly different from control, P < 0.05.