Figure 5. The E/M state mediates invasive and metastatic abilities in DCIS^CAF2cy.

(A) Real-time PCR of the indicated cells measuring ZEB1 expression (left). Immunoblotting of the described cells using anti-ZEB1 and anti–α-tub antibodies (right). (B) Real-time PCR of the indicated cells measuring E-cad expression. (C) Cell invasion evaluated by scratch wound assay (n = 8) using the indicated cells. (D) Cell invasion evaluated by organotypic invasion assay using the indicated cells (n = 3). H&E staining (right-left) of the organotypic gel containing the indicated cancer cells (arrows) and mammary fibroblasts (arrowheads), and its immunofluorescence (right-right) using anti–E-cad and anti-ZEB1 antibodies. The E^hi (simple arrow) and E/M (triangular arrow) cancer cells, and nuclear ZEB1⁺ stromal cells (arrowheads) are shown (right-right). Scale bars, 30 μm (H&E), 10 μm (IF). (E) Lung metastases evaluated by fluorescent intensity at 30 d after intravenous injection of the indicated tdTomato⁺ cells into mice. Data information: Asterisk indicates a significant difference relative to GFP-shRNA–expressing DCIS^CAF2cy (A–E). t test (A–D) and Mann–Whitney U test (E). The horizontal line represents the mean value (E). Error bars, SE. IF, immunofluorescence.

Source data are available for this figure.