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Fig. 3. Cell-mixing experiment. (A-C) After multiple intraperitoneal BrdU
injections, almost all cells derived from En double mutants were positive for
BrdU (B) when attached to a coverslip after 1 hour. (C) Example of
double-labelled TH-positive neurons (arrow) (TH, green; BrdU, red). (D-F) In
the mixture of En double mutant cells and control, no TH-positive,
BrdU-labelled cells were present after 3 days. (G) Number of TH-positive cells
after 24 hours and 72 hours in vitro: control (wild type), mixture of
heterozygote
(En1+/–;En2–/–) and
homozygote (En2–/–). In controls, the number
of mDA neurons decreased only slightly between 24 hours and 72 hours.
Regardless of culture conditions, the numbers of TH-positive cells derived
from engrailed double mutants decreased almost to zero after 72 hours from a
baseline of an average of 45 TH-positive mutant cells at 24 hours.
n=12 for each bar (Student’s t-test,
P<0.001). Error bars indicate s.d. Scale bars: 20 µm.
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