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. Author manuscript; available in PMC: 2021 Apr 7.
Published in final edited form as: Nature. 2020 Oct 7;586(7828):275–280. doi: 10.1038/s41586-020-2776-9

Fig. 2. Retinol metabolism and RXR activity in enterocyte differentiation.

Fig. 2

a, Top, images of organoids treated at day 0 as indicated (DMSO, dimethyl sulfoxide; control; RXRi, treatment with RXR antagonist Cpd2170). Bottom, quantified aldolase B intensity. b, Colour composite images show antibody staining for DLL1 and YAP1 under different treatments at the indicated time points. Scale bars, 20 μm. D, DAPI. Greyscale images show YAP1 localization in the highlighted regions. Scale bars, 10 μm. c, Upper four images show antibody staining for lysozyme, aldolase B and YAP1 under different treatments at day 3. Scale bar, 50 μm. Bottom, YAP1 localization in highlighted regions. Scale bar, 10 μm. d, Retinol metabolism. Cyp26, cytochrome P450; RE, retinyl ester. e, Effect of knocking out Aldhlal on organoid development. Top, images show antibody staining for ALDH1A1; bottom, antibody staining for lysozyme and aldolase B. ALDH1A1i, ALDH1A1 inhibition with A37; pLenti-EF1-EGFP, infection and selection control. f, Left, day 4 organoids under different treatments (ALDH1A1i, day 0; atRA, day 3), in medium with (control) and without vitamin A. Right, quantified aldolase B intensity. Asterisks show statistical significance for the same treatment with/without vitamin A; two-sided t-test; NS, not significant; P values shown in the plot. a-c, e, f, Representative images of organoids (n = 6 independent replicates from n = 3 independent experiments), cultured from single cells. b, Day 2 plus 9 h: organoid middle plane, day 3 and 4: MIP of confocal z-stacks. Nuclei are stained with DAPI; scale bar, 50 μm. a, f, Violin ranges extended by standard deviations; box plots show quartile ranges; whiskers show value intervals; white dots show median values; n values refer to the number of individual organoids in conditions.