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Fig. 3. Cyclopamine reduces Nkx2.1 expression and alters MGE progenitor fate in
vitro. (A-B') Immunofluorescence labeling of re-sectioned (12 µm)
E12.5 telencephalic explants cultured for 24 hours. (A,B) Nkx2.1 and BrdU
labeling of control sections cultured in 0.05% EtOH. (A') Slices
cultured in 5 µM cyclopamine have reduced expression of Nkx2.1 in the
ventricular zone, whereas (B') BrdU incorporation appears unchanged.
(C-F) Immunofluorescence labeling of somatostatin (Som, red), GFP (green) and
BrdU (blue) in cell cultures grown for 14 DIV. The vast majority of
transplanted cells from donors of this age have neuronal morphologies. Arrows
indicate neurons triple-labeled for Som/GFP/BrdU. (G) Overall transplant cell
number is unaffected by cyclopamine. (H) Cyclopamine reduces the percentage of
Som- and Pv-expressing interneurons generated from MGE transplants. (I) The
percentage of surviving, transplanted neurons (GFP+) that incorporated BrdU
prior to transplant is unaffected by cyclopamine after 5 DIV, and slightly
reduced after 14 DIV. (J) Cyclopamine dramatically reduces the number of BrdU+
transplanted cells co-labeling for somatostatin. t-test:
*P<0.05, **P<0.005. Scale bar in A: 200 µm.
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