Fig. 2

Validating Crainbow functionality. a Diagram of NCAT-Crainbow mice. See also Table 1 and Supplementary Fig. 1. b NCAT^VilCre small intestine (N = 10 mice, PND9–PND20) was prepared as a whole-mount and confocal imaged for XFP expression in the intestinal crypts. c Inset in “b” at higher magnification. d Crypts were color segmented and counted. The experimentally observed value was normalized to the predicted stochastic outcome (0.33/XFP). The asterisk denotes statistical significance by one-way ANOVA (TagBFP vs. mTFP1: p < 1e–6, TagBFP vs. mKO: p < 1e–6, mTFP1 vs. mKO: not significant). NCAT^CreER/T2 mice (N = 8, 19–22 weeks of age) were injected with 200 mg/kg tamoxifen. Mice were sacrificed 12 days post tamoxifen injection, and the small intestine was vibratome sectioned and antibody stained to recover the quenched TagBFP signal. e Sections were imaged by confocal microscopy for XFP expression (TagBFP: cyan, EYFP: mTFP1, mKO: magenta; segmented nuclear masks shown and overlaid with surface rendered tissue outline). f Rotated inset in “e” at higher power. g Nuclei were segmented, counted, and normalized to the predicted stochastic outcome (0.33/XFP). The asterisk denotes statistical significance by one-way ANOVA (TagBFP vs. mTFP1: p < 1e–6, TagBFP vs. mKO: p < 1e–6, mTFP1 vs. mKO: 8.4e–4). (SEM included for each graph). Scale Bars = 100 µm in b, c, f and 2 mm in e. Source data are provided as a “Source Data file”.