HMVECs transfected with Akt dominant-negative mutants or Src antisense ODNs show significantly less sE-selectin–induced tube formation.

HMVECs were transiently transfected with a pcDNA3 plasmid containing ERK1/2 and Akt dominant-negative mutants or with Src sense or antisense ODN. Plasmid with no insert was used as a negative control. HMVECs transfected with mutants were used for Matrigel in vitro HMVEC tube formation assays 72 hours after transfection, whereas HMVECs transfected with Src ODNs were used 16 hours after transfection. (A) Photomicrographs of a representative assay for (i) HMVECs transfected with no insert, (ii) HMVECs transfected with no insert and stimulated with sE-selectin, (iii) HMVECs transfected with an Akt mutant, (iv) HMVECs transfected with an ERK1/2 mutant, (v) HMVECs transfected with Src antisense ODNs and stimulated with sE-selectin, and (vi) HMVECs transfected with Src sense ODNs. Original magnification, × 20. (B) Quantitative data for HMVEC tube formation are expressed as number of tubes/well ± SEM from 4 independent experiments. *Represents a significant difference (P < .05) between groups. HMVECs transfected with Akt dominant-negative mutants and Src antisense ODNs showed significantly less tube formation compared with HMVECs transfected with no-insert plasmid.