Figure 3
PLVAP expression is regulated by eNOS in HDMVECs and HUVECs, but not in HRECs. (a) PLVAP mRNA levels in VEGF-A-stimulated (25 ng/mL for 48 h) siNT- and siNOS3-treated HDMVECs, HRECs and HUVECs in n = 3–4 independent experiments. (b) Representative confocal images of PLVAP immunofluorescence staining in siNT- and siNOS3-treated HDMVECs and HRECs. Cells were cultured in the presence or absence of VEGF-A (25 ng/mL for 48 h). Scale bars: 20 µm. (c) Quantification of PLVAP immunofluorescence staining data shown in (b), n = 3 independent experiments. (d) PLVAP mRNA levels in control, L-NAME pretreated (100 µM, 30 min), VEGF-A-stimulated (25 ng/mL for 48 h) or a combination of L-NAME pretreated and VEGF-A-stimulated HDMVECs, HRECs, HUVECs and BRECs, in n = 3–4 independent experiments. (e) PLVAP mRNA levels in HDMVECs, HRECs and HUVECs after repeated treatment with NO donor SNAP (50 µM, 100 µM or 200 µM) over a 48 h incubation period (cells were stimulated 3 times for 4 h per day with freshly prepared SNAP), in n = 3–4 independent experiments. Data are represented as mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001; ns, not significant (one-sample t-test or Student’s t-test).
