Fig. 3: Ductal and vascular consequences resulting from specific breast cancer mutations.
a Left: Cartoon of co-culture paracrine experimental setup with mosaic mutant/wild-type ducts. Ducts were generated from MCF10A cells stably expressing empty vector (EV), wild-type hemagglutinin (HA)-tagged ErbB2 (ErbB2amp), or constitutively active HA-tagged PI3Kα H1047R (PI3KαH1047R) along with each associated vascular compartment. Right: Western blot of lysates from EV, ErbB2amp, or PI3KαH1047R MCF10A. b Composite stitched maximum intensity projection micrographs of endothelial vessels co-cultured for five days along with their corresponding modified ducts. Endothelial vessels: phalloidin (green), DAPI (blue), epithelial ducts: phalloidin (cyan), DAPI (blue). c Diffusive permeability (PD) of 70 kDa dextran across the endothelial barrier (n = 8, 8, 6 vessels examined across three independent experiments; one-way ANOVA with Bonferroni post test, **P = 0.0049, *P = 0.012) and (d) endothelial vessel diameter in each co-culture setting (n = 8, 8, 6 vessels examined across three independent experiments; one-way ANOVA with Bonferroni post test). e Left: Quantification of cortical actin measured from phalloidin labeled micrographs (n = 12, 13, 13 cells taken from three independent vessel experiments one-way ANOVA with Bonferroni post test, ****P = 7.6845e−06, ***P = 0.0003). Right: Averaged, cross-sectional actin distribution in endothelial vessels co-cultured with specific mutant ducts (n = 11 cell actin profiles taken from three independent vessel experiments). 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.
