Involvement of Notch Signaling in Wound Healing
Figure 4
Notch signaling regulates the motility and proliferation of vascular endothelial cells.
a and b. Cultured human vascular endothelial cells (HUVEC) were treated with either vehicle (Control), g-secretase inhibitor (GSI; 10 µM DAPT) or Jagged peptide (15 µM) in conditioned medium and plated into chemo-attractant medium consisting regular growth medium with 10% FBS and other growth factors and cell migration was evaluated using a 24 well Transwell chamber assay. Representative images are shown in panel and a and quantitative data are shown in panel b (values are the mean±SEM for cells per 100×field; n = 3-4). *p<0.001 compared to control values. #p<0.001 compared to SCP values. c–f. HUVEC monolayers were mechanically wounded with the sterile tip of a 20–200 µl pipette tip following treatment with vehicle, GSI (10 µM DAPT), scrambled control peptide (SCP; 15 µM) or Jagged peptide (15 µM). Representative images are shown in c and e, and quantitative data on cell migration in d and f. Values are the mean and SEM (n = 3 separate experiments). *p<0.001. g and h. HUVEC were seeded on Matrigel-precoated wells and cultured in the presence of low-serum medium with GSI (10 µM DAPT), SCP (15 µM) or Jagged peptide (15 µM). Tube formation, designated as the number of branch points/100×field) was evaluated 18h after cell plating. Representative images are shown in g and quantitative data in h. Values are the mean and SEM (n = 12–16 cultures). *p<0.001. i. Cultured HUVEC were treated with GSI (10 µM DAPT), scrambled peptide (SCP; 15 µM) or Jagged peptide (15 µM) for the indicated time periods and relative cell numbers were estimated (n = 4–6 experiments). *p<0.001, #p<0.01 compared to the control cultures.