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Fig. 4. Apoptosis and proliferation of ENCCs. (A,B) Activated
caspase 3 (red) and HuD (green) double immunostaining on E13.5 midgut
sections. No apoptotic neuron is detected in mutant and control. White
arrowheads indicate autofluorescent red blood cells in the submucosal zone.
The red arrowhead shows an activated-caspase-3+ HuD-
cell in the mesentery. (C,D) PCNA immunostaining on E12.5
X-Gal-stained gut sections. (F,G) BrdU (red) and Phox2b (green) double
immunostaining on E12.5 gut sections. (E,H) Percentages of
ß-gal+ cells that were PCNA+ (E) and percentages of
Phox2b+ cells that were BrdU+ (H), in control and mutant
midgut and caecum (cell counts from several preparations were pooled; n, total
number of ENCCs examined; 
2 test, P>0.05). Scale
bar: 100 µm in A,B; 20 µm in C,D,F,G.
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