Fig. 2: Expression of various markers in 3D-UHU-TU.

a E- and N-Cadherin (grey) in 3D-UHU-TU in 100% urine environment 2 days post-implantation (3D-UHU layer below plane of focus). For merged images, Cadherin = Red, nuclei = cyan. Scale bars = 50 µm. b H&E and IHC in models maintained for 6 days in 100% urine environment. Images are representative of 3 separate biological replicates. Bars = 50 µm. c CK20 expression (left panels; greyscale) with 3D merges. Models were stained for F-Actin (magenta), which highlights the actin cytoskeleton, useful for providing an overview of cell morphology, nuclei (cyan), cancer spheroid (green) and CK20 (yellow). CK20 on urothelium are highlighted with white arrows. Models maintained for 6 days in 100% urine environment. d Migration phenotypes magnified from Fig. S2B, C. MMP-2 & -9 (red) and nuclei (cyan) stained. Scale bars = 10 µm. Representative images from 3 independent experiments.