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. 2015 Aug 12;34(20):2522–2536. doi: 10.15252/embj.201490583

Figure EV3.

Figure EV3

Chronic Notch activation in LGR5+ stem cells does not induce intestinal polyps
  • A, B Goblet cells in NICD+ ileal crypts were analyzed by co-staining for GFP (cytoplasmic, Lgr5; nuclear, NICD) and Alcian Blue in control (n = 3) or Lgr5; ROSANICD (n = 3) mice 1 month post-TX. Nuclear GFP was observed in a patchy pattern consistent with the known mosaic expression pattern of Lgr5-EGFP-IRES-CreERT2. Recombined crypts with NICD-GFP expression (asterisk) in Lgr5; ROSANICD mice exhibited a reduction in goblet cells, in accordance with the known effect of Notch signaling to block secretory cell differentiation. Arrowheads indicate LGR5-GFP+ intestinal stem cells at the base of the crypts. Scale bars: 50 μm.
  • C–J No polyps were observed in intestine 6 months after TX induction of NICD. H&E analysis of Lgr5 control (n = 7) (C–F) and Lgr5; ROSANICD (n = 7). (G–J) small intestine (C–E; G–I) and colon (F, J). Scale bars: 100 μm.
  • K, L Proliferation was measured by EdU incorporation and found to be unchanged in Lgr5; ROSANICD mice. Scale bars: 50 μm.
  • M, N qRT–PCR analysis of Olfm4 and Hes1 in Lgr5 and Lgr5; ROSANICD intestine revealed no change to stem cell marker or Notch target gene expression. Data are expressed as mean ± SEM (n = 7 mice per group).