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. Author manuscript; available in PMC: 2017 Nov 3.
Published in final edited form as: Nature. 2017 May 3;545(7653):238–242. doi: 10.1038/nature22313

Figure 2. Lineage tracing of Lgr5-eGFP+ ISCs upon systemic Rspo inhibition versus overexpression.

Figure 2

a, Ad LGR5, Rnf43 or Znrf3 ECDs ablate Lgr5-eGFP reporter signal and induce precocious Lgr5+ ISC villus lineage tracing Lgr5-eGFP-IRES-CreER; Rosa26-tdTomato mice, d2 after adenovirus and tamoxifen. b, Multi-lineage differentiation in d7 LGR5 ECD-induced tdTomato+ lineage stripes. c, RNA-seq heatmap of reciprocal changes in FACS-sorted Lgr5-eGFP+ cells, d1.5 after Ad Rspo1 or Ad LGR5 ECD, Lgr5-eGFP-IRES-CreER mice. d, Ad Rspo1 or Ad RSPO2 both expand Lgr5+ ISCs, whose lineage traces are profoundly crypt-confined with striking repression of red progeny and villus lineage tracing. In contrast, Ad LGR5 ECD ablates Lgr5-eGFP expression and drives premature Lgr5+ ISC lineage tracing. Lgr5-eGFP-IRES-CreER; Rosa26-tdTomato duodenum, d2–7. Bars = 50 μm. In a–d, tamoxifen was given at day 0. All experiments used n=3 mice per condition and were repeated at least twice, except for c, which used n=2–3 mice per condition and performed once.