Figure 7.

ER stress-generated MPs impaired angiogenic capacity in HUVECs. (A), HUVECs were incubated in complete M200 medium (CTL or no MPs), or in complete culture medium after treatment with CTL MPs, TG MPs, TG+PBA MPs or PBA MPs (20 μg/ml for 48 h). Angiogenic capacity of HUVECs was assessed by Matrigel-based tube formation assay. Images are representative of three independent experiments; images were taken after 4 h. To quantify angiogenesis, the length of tubes formed was counted in five random fields for each well using WimTube software from Wimasis Image Analysis. Bars represent the pooled data expressed as the total length of tubes formed in each condition. (B,C), relative mRNA expression levels of angiogenic factors VEGF-A (B) and FGF-2 (C) after treatment with MPs (20 μg/ml for 48 h), normalized against housekeeping gene β-actin (n = 3 per group). Data are presented as mean ± SEM and were analyzed by one-way ANOVA followed with Tukey's multiple comparison test. *P < 0.05, **P < 0.01 vs. CTL (No MPs) or vs. indicated groups; ##P < 0.01, ###P < 0.001 vs. CTL MPs.