Fig 2: HG-SOC invasion is regulated by ET-1/Intß1 signaling.A si-SCR, or si-TLN1 or si-ARRB1 OVCAR3 cells treated with ET-1 and/or AMB and/or ATN161 were allowed to invade fibronectin/type I collagen plugs in an inverted invasion assay (48 h). Cells were stained with PKH67, and serial optical sections (10 µm intervals) were acquired. The invasion was measured by dividing the sum of signal intensity of all slides beyond 20 µm (invading cells) by the sum of the intensity of all slides (total cells). n = 2, one-way ANOVA, Tukey post hoc analysis. Scale bar, 200 µm. B SKOV3 cells plated on a monolayer of MCs grown on fibronectin/type I collagen in a polystyrene scaffold were allowed to invade for 7 days in the absence and presence of ET-1 and/or AMB and/or ATN161, then fixed with Bouin’s solution and paraffin-embedded scaffolds were then cut into thin slices (10 µm). The images show cell invasion in a 3D organotypic model. Hematoxylin and eosin staining are shown. C Sections as in (B) were stained for active Intß1 (green) and DAPI (blue) detection. The corresponding transmitted light images are also shown. Arrows depict the top side of the scaffold where cells were plated. Scale bar, 50 µm.