Figure 4 from Novel Syngeneic Cell Lines for Studying High-Risk BRAFV600E-Driven Colorectal Cancer In Vivo

All NaJa cell lines successfully engraft into syngeneic immunocompetent C57BL/6N mice and recapitulate their individual 2D morphologies in vivo. A, Schematic overview of the PVI of NaJa cells (n = 2 NaJa-D, n = 10 NaJa-F, and n = 3 NaJa-G). B, Left, livers showing metastatic nodules 4 weeks after injection (black arrows) and invasive growth in the pancreatic tissue (blue circles). Scale bar, 1 cm. Hematoxylin and eosin: Histology showing that all NaJa cell lines establish tumors (pale blue) in the liver parenchyma (pink) with a differentiation state matching their epithelial or mesenchymal in vitro morphology. Right, IHC and immunofluorescence (IF) staining of proliferation marker Ki-67 (left) and intestinal differentiation markers CDX2 (middle) and E-cadherin (right). L, liver; T, tumor. Scale bar (overview), 500 μm, scale bar (zoom, IHC, and IF), 50 μm. C, Quantification of Ki-67 and CDX2 IHC staining in NaJa cell–induced liver metastases. Each dot represents an individual tumor; different shapes indicate individual tissue sections, and colors denote individual mice. Data are shown as the median ±95% confidence interval (CI).Statistical significance was assessed using the Kruskal–Wallis test followed by Dunn multiple comparison test. **, P < 0.01; ****, P < 0.0001. D, IHC staining of ERK and its downstream target DUSP6. Red arrows indicate examples of nuclear ERK staining. L, liver; T, tumor. Scale bar, 50 μm. [A, Created by J. Traichel in BioRender. Brummer, T. (2025) https://BioRender.com/8ogg64c.]