Figure 1.

2-DG (mM) depletes intracellular ATP and inhibits in vitro capillary-like structure formation in HBMEC. In order to assess the impact of ATP requirement for in vitro tubulogenesis, HBMEC were seeded on top of Matrigel as described in the Methods section and left to form structures. A) Capillaries were then treated with 2-DG (mM) or Mannose (mM). B) The impact of 2-DG was also monitored during the formation of the structures, where cells were treated with various concentrations of 2-DG 30 min after seeding of the cells on top of Matrigel, and structure formation monitored after 18 h. Representative phase contrast pictures were taken. C) The extent of three-dimensional capillary-like structure formation (tubulogenesis) and of cell survival was assessed as described in the Methods section. D) Intracellular ATP as well as total protein content were assessed as described in the Methods section in vehicle- or 2-DG-treated cells. The length of the tube network was quantitated using Northern Eclipse software.

Notes: Values are means of two independent experiments (*P < 0.01 versus control alone); bars, ± SD.

Abbreviations: HBMEC human brain microvasular endothelial cell; ATP, adenosine triphosphate; 2-DG, 2 deopy-D-glucose.