Figure 3. Apc-tumor model at single-cell resolution using the MADM-18 line.
(A) Schematic representation of MADM labeling (green, GFP; red, tdT) and respective cellular genotypes in wild-type MADM-18GT/TG;HprtCre/+ mice.
(B and C) P-H3 staining (white) in small intestine in MADM-18GT/TG;HprtCre/+ mice at 3 months of age. (B) Overview of unicolor (monoclonal) green wild-type crypt-villus units with insets highlighting non-proliferative villus epithelium (i) and a proliferative cell within the crypt (white arrow) (ii); (C) overview and unicolor (monoclonal) red wild-type crypt-villus units with insets highlighting non-proliferative villus epithelium (iii) and a proliferating cell within the crypt (iv).
(D) Quantification of the percentage of intestinal structures displaying MADM labeling. Data obtained from n = 3 male MADM-18GT/TG;HprtCre/+ mice at 3 months of age.
(E) Schematic representation of MADM labeling (green, GFP; red, tdT) and respective cellular genotypes in genetic mosaic MADM-18GT/TG,Apc;HprtCre/+ mice. (F) P-H3 staining (white) in small intestine in male MADM-18GT/TG,Apc;HprtCre/+ mice at 3 months of age with insets highlighting a proliferating adenoma cell at the boundary to the non-proliferative villus epithelium (white arrow) (i), proliferating adenoma cells within the tumor (white arrows) (ii), non-proliferative normal villus epithelium (iii), and proliferative cells within a normal crypt compartment (iv).
(G) Quantification of the percentage of intestinal structures displaying MADM labeling. Green Apc−/− cells display 100% transformation and tumor initiation, whereas red wild-type cells solely give rise to normal crypt-villus-units. Data obtained from n = 3 male MADM-18GT/TG,Apc;HprtCre/+ mice at 3 months of age.
(H and I) Summary of MADM labeling in small intestine of control MADM-18GT/TG;HprtCre/+ (H) and genetic mosaic MADM-18GT/TG,Apc;HprtCre/+ mice (I). Note that in the mosaic, red wild-type cells give rise to normal crypt-villus units and green Apc−/− cells initiate tumor development and subepithelial invasion of adenomas. Nuclei were stained using DAPI. Scale bar: 100 μm (B, C, and F) and 25 μm (i–iv).
