Fig. 2: Vascular morphogenesis driven by mammary paracrine signaling.

a Cartoon of co-culture paracrine experimental setup. MCF10A cells expressing IRES-GFP or VEGF-IRES-GFP were seeded into the duct channels adjacent to an endothelialized vessel. Phase-contrast images of vessels with control (left) and VEGF expressing ducts (right) after two days in co-culture. b Quantification of endothelial vessel diameter after three days of culture with either GFP or VEGF expressing MCF10A cells (n = 6, 9 vessels examined across three independent experiments; two-tailed, unpaired Student’s t test **p = 0.0019). c Quantification of sprout number per endothelial vessel after three days of culture with acellular basal assay medium (BM), GFP expressing ducts (GFP), or VEGF expressing ducts in which DMSO (VEGF + Veh) or 10 µM Semaxanib (VEGF + Sem) was delivered to the vasculature. (n = 9, 6, 9, 8 vessels examined across three independent experiments; one-way ANOVA with Bonferroni post test ***P = 0.000014, **P = 0.0011). d Maximum intensity projection micrographs of tissue base to medial section of endothelial vessels co-cultured with GFP or VEGF ducts and treated with DMSO or 10 µM Semaxanib; phalloidin (green) and DAPI (blue). Arrows indicate nascent angiogenic sprouts. For all plots, values mean ± s.e.m and all images are representative of at least three independent experiments. Source Data are provided as a Source Data file.