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

Figure 2.

Figure 2

Notch regulates stem cell function
  • A–D Gastric stem cell proliferation was measured by morphometric analysis of GFP/Ki67 cell numbers in Notch-inhibited or Notch-activated antral tissue, showing decreased LGR5-GFP stem cell proliferation after DBZ treatment (A, n = 5) and increased stem cell proliferation in Lgr5; ROSANICD mice (C, n = 4) 1 month post-TX treatment. *P = 0.046 and **P = 0.002 versus vehicle using Student's t-test. (B, D) Olfm4 gene expression was measured by qRT–PCR in antral RNA from vehicle (n = 10) versus DBZ (n = 9) (*P = 0.033 using Student's t-test) and vehicle (n = 3) versus Lgr5; ROSANICD mice (n = 3) (*P = 0.026 using Student's t-test) 1 month post-TX treatment. Data are expressed as mean ± SE.
  • E–H Antral organoids were established from gastric glands isolated from mice treated with vehicle or DBZ, or from control (Lgr5) or Lgr5; ROSANICD mice 1 week post-TX. Scale bars: 200 μm.
  • I–L Plating efficiency of antral organoids established from (I) vehicle (n = 3) or DBZ-treated (n = 3) mice (*P = 0.046 using Student's t-test) or (K) Lgr5 (n = 3) or Lgr5; ROSANICD mice (n = 3) 1 week post-TX (*P = 0.012 using Student's t-test). The number of organoids was counted 48 h post-plating, and normalized to the number of glands initially plated. (J, L) Organoid size was measured 4 days post-plating. (L) *P = 0.025 versus Lgr5 using Student's t-test. Data are expressed as mean ± SE.
  • M–O Single Lgr5-GFP stem cells were isolated by FACS from vehicle (n = 4) or DBZ-treated (n = 4) mice. See Appendix Fig S4 for FACS gating scheme. (O) Organoid establishment efficiency was quantified 3 days post-plating. *P = 0.014 versus vehicle using Student's t-test. Data are expressed as mean ± SE.