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Fig. 2. Effect of ectopic Notch activation on endocrine and exocrine
differentiation in explant cultures of E10.5 dorsal pancreatic buds. E10.5
dorsal epithelial buds were infected with lentiviral vectors encoding either
GFP alone, GFP and Hes1, or GFP and Notch1-IC, then recombined with mesenchyme
and allowed to differentiate in vitro. (A-C) Examination of intact buds
demonstrates mosaic GFP expression on day 1 following lentiviral infection.
(D-O) Deconvolution fluorescent microscopy demonstrating expression of
endocrine and exocrine markers in infected E-cadherin-positive epithelial
cells marked by GFP expression, assessed on day 6 following lentiviral
infection. Cells infected with GFP alone are able to complete either endocrine
or exocrine differentiation programs, as assessed by co-expression of GFP with
insulin (D), glucagon (G), amylase (J), or Ptf1-P48 (M). Cells infected with
GFP in combination with either Hes1 or Notch1-IC maintain the capacity for
alpha-cell differentiation, as evidenced by co-expression of GFP and glucagon
(H,I), but are unable to undergo beta-cell differentiation, indicated by
absence of cells co-expressing GFP and insulin (E,F). Cells infected with GFP
in combination with either Hes1 or Notch1-IC are also unable to undergo acinar
cell differentiation, as evidenced by absence of cells co-expressing GFP and
amylase (K-L). In contrast, expression of Ptf1-P48 is preserved in cells
expressing GFP in combination with either Hes1 or Notch1-IC (N,O). Arrowheads
in D,G,H-J,M-O indicate examples of individual cells co-expressing GFP and
indicated marker. Scale bars: 10 µm.
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