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. 2018 Sep 4;4:49. doi: 10.1038/s41421-018-0051-0

Fig. 2. CHIR and LDN together maintain Lgr5+ ISCs.

Fig. 2

a Representative images and quantitation of colony numbers of 5,000 FACS-isolated Lgr5-GFP cells cultured with ENR plus Wnt3a (ENRW), CHIR and LDN (2ki) or ENR plus CHIR (ENRC) for 7 days. b, c GFP fluorescence images (b) and FACS analysis of GFP expression (c) of organoids derived from single small intestinal Lgr5-GFP cells cultured with ENRW, 2ki and ENRC for 7 days. d–f Representative bright-field (d), GFP fluorescence images (e), and FACS analysis of GFP expression (f) of small intestinal organoids cultured with ENR and 2ki for 2 months. g Scatter plots comparing the gene expression between ENR and 2ki organoids (spearman correlation is 0.976). The Lgr5+ ISC signature genes are showed in blue dots. h Heat maps showing the RNA expression of Lgr5+ ISC signature genes. i Lgr5-GFP cells sorted from organoids cultured with ENR or 2ki for 2 months were shifted to ENRW culture for 7 days, and then representative images and quantitation of colony numbers were shown. Scale bars, 100 μm. ***P < 0.001, **P < 0.01, and *P < 0.05 analyzed by two-way ANOVA test. Error bars, s.d. n = 4 wells from 2 mice in c and f, n = 9 wells from 3 mice in a, and n = 6 wells from 2 mice in i. Refer to Supplementary Figure 3 for detailed analysis