Figure 3.
ACF has anti-proliferative, pro-apoptotic, and anti-angiogenic effects on human endothelial cells
(A) Percentage of HUVECs surviving different concentrations of ACF (0.1–100 μM, 16 h) or CPT (10 μM, 16 h). DMSO served as a negative control (CTL). n = 4, one-way ANOVA with Dunnett post hoc test. (B) Calculation of IC50, indicated by the percentage of HUVECs surviving different concentrations of ACF (−7 to −4 log mol/L, 16 h). n = 7, nonlinear regression model. (C) Caspase-3/7 activity assay of HUVECs treated with or without ACF (10 μM, 16 h) or CPT (10 μM, 16 h). n = 6, one-way ANOVA with Bonferroni post hoc test. (D) Relative wound closure of HUVECs treated with or without ACF (16 h, 10 μM). DMSO served as a negative CTL. n = 24, two-way ANOVA with Bonferroni post hoc test. (E) Matrigel assay of HUVECs treated with or without ACF (10 μM, 16 h). DMSO served as a negative CTL. n = 3, paired t test. (F–H) Spheroid outgrowth assay (F) and quantification of sprout numbers (G) and cumulative sprout lengths (H) of HUVECs treated with or without ACF (10 μM, 16 h). Cells were studied under basal conditions or after treatment with VEGF-A (30 ng/mL). The scale bar indicates 100 μm. n = 30. One-way ANOVA with Bonferroni post hoc test. Data are mean ± SEM. ∗p < 0.05.