Figure 3.

Role of H₂S production on in vitro angiogenesis in response to zofenoprilat. ECs were incubated with zofenoprilat (10 μM, 40 min or 5 days, respectively), pretreated or not with PAG (3 mM, 30 min), and adhesion (A) or proliferation (B) were assessed. Data represent counted cells per well. n = 3. ^*P < 0.05, ^**P < 0.01 versus untreated cells. ^#P < 0.05, ^##P < 0.01 versus zofenoprilat-treated cells. Cell migration (C) was evaluated using scratch wound healing assay on ECs treated with 0.1% FBS (a), zofenoprilat (10 μM, 18 h, b), PAG (3 mM, 30 min pretreatment, c) or zofenoprilat with PAG (d). Representative pictures of wounded EC monolayers are shown. n = 3. Quantification (e) of cell migration presented as % area of wound. ^***P < 0.001 versus untreated cells; ^##P < 0.01 versus zofenoprilat. (D) Pseudocapillary formation in Matrigel by ECs exposed to 0.1% FBS (a), zofenoprilat (10 μM, b), PAG (3 mM, 30 min pretreatment, c) and PAG + zofenoprilat (d) after 18 h. Cells were labelled with DY554-phalloidin, and quantification (e) of pseudocapillaries was performed by counting the numbers of complete circles per well. n = 3. ^**P < 0.01 versus untreated cells; ^##P < 0.01 versus zofenoprilat.