Fig. 2

In the presence of VEGF, IL-11 cis-signaling has anti-angiogenic effects on vascular endothelial cells while trans-signaling enhances their pro-angiogenic and pro-migratory potential associated with an enhanced respiratory rate. (A) Spheroid sprouting assay with HUVECs exposed to endothelial basal medium (EBM = negative control), VEGF (positive control), IL-11 + VEGF or IL-11 + sIL-11Rα+ VEGF for 17 h. N = 3 independent experiments with 15-22 spheroids per group and experiment. Statistical test: Kruskal-Wallis Test adjusted for multiple testing, *p<0.01. Relative sprouting length (RSL), scale bar 20 µM. (B) Spheroid sprouting assay with HRMVECs exposed to VEGF (positive control), IL-11 + VEGF or IL-11 + sIL-11Rα + VEGF for 17 h. N = 2 independent experiments with 10-22 spheroids per group and experiment. Statistical test: Kruskal-Wallis Test adjusted for multiple testing, *p<0.01. Relative sprouting length (RSL), scale bar 20 µM. (C) Scratch wound assay using HUVECs: Migratory effect of IL-11 + sIL-11Rα + VEGF and IL-11 + VEGF on HUVECs over 24 h. N = 3 independent experiments each including 6-8 technical replicates. Relative Wound Density (RWD). Exemplary analysis of the conditions 11 h after stimulation, statistical test: Kruskal-Wallis Test adjusted for multiple testing, *p<0.01. Scale bar 200 µM. (D) Representative graph of the oxygen consumption rate (OCR) of HUVECs after pretreatment with above mentioned cytokines for 15 h. N = 3 with each 4-8 technical replicates. Statistical test: Kruskal-Wallis Test adjusted for multiple testing, *p<0.05. (E) Representative graph of the normalized extracellular acidification rate (ECAR) of HUVECs after pretreatment with above mentioned cytokines for 15 h. N = 3 with each 4-8 technical replicates. Statistical test: Friedman Test adjusted for multiple testing, *p<0.05