HMCLs up-regulate Tie2 expression by endothelial cells. Human BM endothelial cell line BMEC-60 (2 × 106) was cocultured in the presence or absence of a transwell system with HMCL (5 × 106) XG-6 in 6-well collagen-coated plates. After 24 hours, mRNA was extracted from BMEC-60 alone, in the control and transwell condition, or from pooled BMEC-60 and XG-6 in cell-contact condition. (A) Ang-1, Ang-2, and Tie2 expression were evaluated by RT-PCR, and ET-1 expression was used to document that an equal amount of endothelial cells was analyzed. (B) Ang-1, Ang-2, and Tie2 proteins were assessed by Western blot analysis after 48 hours. Figures are representative of 3 independent experiments (C indicates BMEC-60 control; transW, transwell; C+XG-6, contact coculture).
