Fig. 3
From: A cancer rainbow mouse for visualizing the functional genomics of oncogenic clonal expansion
Widespread expansion of oncogenic clones during perinatal development. a Diagram of MCAT-Crainbow mice. See also Table 1 and Supplementary Fig. 5. b MCATVilCre small intestine (N = 10 mice, 3–6 weeks of age) prepared as a wholemount and confocal imaged. c Inset in “b” at higher magnification. d MCATVilCre Crypts were color segmented, counted and normalized to the positional bias calculated in NCATVilCre mice. Asterisk denotes statistical significance by one-way ANOVA (mTFP1 vs. EYFP: p = 0.003, mTFP1 vs. mKO: p = 0.016, EYFP vs. mKO = 3e–6). e Immunostaining for FLAG, V5, or HA epitopes (magenta) specific to each ßcat isoform in MCATVilCre small intestine vibratome slices and merged with fluorescent lineage markers (mTFP1: cyan, EYFP: yellow, and mKO: orange). Arrows denote isoform expression with cognate lineage reporter (FLAG and mTFP1, V5 and EYFP, and HA and mKO). Corresponding insets depict higher magnification images. Arrowheads denote membrane-localized ßcat, whereas asterisk denotes nuclear-localized ßcat. Epitope stains (magenta) are also presented as merged and as a single-channel image with its cognate fluorescent lineage reporter (green). f HEK cells were transiently transfected with MCAT isoforms, fixed, stained, and imaged for the indicated epitope (magenta) and fluorescent reporter (green). Cells were also cotransfected with epithelial cadherin (CDH1) as indicated. Arrows denote sequestration of ßcat at the plasma membrane, and the asterisk denotes nuclear ßcat. g Wnt signalling activity for each oncogene in the absence of CDH1 (solid bar) or in the presence of overexpressed CDH1 (hatched bar) (N = 6 wells per condition and independently repeated in four experiments). TOP FLASH activity was normalized to WNT/RSPO-stimulated control cells (dashed line). Asterisk denotes statistical significance by two-way ANOVA and Bonferroni’s multiple comparisons test (cyan < 1e–6, yellow = 0.01, magenta = 0.02). (SEM included for each graph). Scale Bars = 1 mm in b, 100 µm in c/e, 15 µm in e: insets 1–3, and 10 µm in f. Source data are provided as a “Source Data file”.
