Fig. 5
From: A cancer rainbow mouse for visualizing the functional genomics of oncogenic clonal expansion
Ectopic expansion of the intestinal stem cell compartment by expression of oncogenic RSPO3. a Multiprobe RNA FISH and confocal microscopy for Top2a (cyan), Lgr5 (yellow), and Rspo3 (magenta) in paraffin-embedded sections of the small intestine (SI) and colon (Col). b ROBOVilCre mice expressing oncogenic human RSPO3. Position 0: control and membrane-targeted FAP-Mars1, Position 1: mTFP1 (cyan) coexpressed with 3×FLAG-RSPO3, EYFP (yellow) coexpressed with V5-PTPRKe1:RSPO3e2–5, and Position 3: mKO (magenta) coexpressed with 3×HA-PTPRKe1–7:RSPO3e2–5 (see also Supplementary Fig. 7). c Gross examination of the ROBOVilCre mice and gastrointestinal tracts at PND16 compared with WT littermate controls. WT and ROBO d small intestines (SI) and e colons (Col) were paraffin embedded and sectioned for H&E pathology and coregistry of indicated cell type markers by multiprobe RNA FISH. Images are insets from whole-slide imaged Swiss rolls (see Supplementary Figs. 8–13). f MCATVilCre and ROBOVilCre mice were injected with EdU (magenta) and coimaged for each lineage reporter by confocal microscopy (mTFP1: cyan, EYFP: yellow, mKO: orange). g ROBOVilCre organoids can grow without exogenous RSPO3 (ENY) and were imaged by confocal microscopy for each lineage reporter. Scale bars = 10 µm in a, 2 cm in c, 100 µm in d, e, 50 µm in f, and 1 mm in g. Source data are provided as a “Source Data file”.
