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Fig. 5. Expression and activity of Ascl3 and Ascl5. (A) Expression of
Aslc3 and Ascl5 in the spinal cord at E10.5 and E16.5. mRNA expression was
detected by RT-PCR using reverse-transcribed (RT+) and non-transcribed (RT-)
RNA samples as templates. Glyceraldehyde-3-phosphate dehydrogenase (Gapdh) was
used as internal control. (B-D) Expression of Ascl1 protein, and of
Ascl3 and Ascl5 mRNA in the VZ of the mouse spinal cord at E10.5. The neural
tube is outlined in C,D. Brackets indicate the position of the
Olig2+ motoneuron progenitor domain (pMN). The horizontal lines
indicate the boundary of the dorsal and ventral aspects of the VZ.
(E-G) Neurogenic and oligodendrogenic activity of Ascl3 and Ascl5.
Neurospheres derived from E13.5 spinal cords were infected with GFP
retroviruses expressing Ascl1, Ascl3 and Ascl5, and subsequently induced to
differentiate for 4 (E) or 10 (F,G) days. The percentages of GFP+
cells that differentiated into TuJ1+ neurons, O4+
oligodendrocytes and GFAP+ astrocytes were quantified
(mean±s.d., three independent experiments).
*P<0.01 compared with control virus-infected culture.
Scale bar: 100 µm.
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