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. 2019 Oct 30;23(2):159–177. doi: 10.1007/s10456-019-09688-8

Fig. 3.

Fig. 3

Fibroblast-derived WNT2 induces vessel growth and sprouting in a 3D angiogenesis co-culture assay. a Skin fibroblasts ectopically expressing WNT2 (BJ1WNT2, red) or with parental BJ1 (gray) were co-cultivated with HUVEC-coated microcarrier beads. WNT2 overexpression was evaluated by RT-qPCR. b After 14 days of co-culture, endothelial structures were stained with CD31 and representative images are depicted. The position of the bead is indicated by a green dotted line. c Image processing was used to quantify vessel areas, sprout numbers, branch points, and sprout length per bead [40]. Blue horizontal lines indicate the mean, error bars are SEM, endothelial structures derived from 40 beads were analyzed for each condition, and P values are indicated. d CAFs (CAF#1) endogenously expressing WNT2 (CAF-NTC, gray) or with a WNT2 knockdown (CAF-siWNT2, blue) were co-cultivated with HUVEC-coated microcarrier beads. WNT2 depletion was evaluated by RT-qPCR. e After 14 days of co-culture, CD31+ endothelial structures were evaluated and representative images are depicted. The position of the bead is indicated by a green dotted line. f Vessel areas, sprout numbers, branch points, and sprout length per bead were measured. Red horizontal lines indicate the mean; error bars are SEM; CAF-NTC, n = 45; CAF-siWNT2, n  = 63; P values are given.