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Fig. 7. (A) Tenascin C (TNC) gene expression in neurospheres from P0 wild-type (WT)
and P0 TNC null mice (K/O) grown for seven days in either epidermal growth
factor (EGF) or fibroblast growth factor (FGF2) (20 ng/ml) assessed by RT-PCR.
Note the band corresponding to TNC mRNA in WT but not null neurospheres.
ß-actin mRNA is detected as a loading control (ß-a), and sizes shown
on the right are in bp. (B-E) Six days after growth factor withdrawal,
neurosphere cells differentiate into oligodendrocytes, as shown by Gal-C
staining in red (B, WT cells; C, TNC null cells), and neurones, as shown by
ßIII-tubulin staining in green (D, WT cells; E, TNC null cells). The
increased number of neurones in the absence of TNC can be seen by comparing D
(WT) and E (TNC null), and is quantified as a percentage of the total cell
numbers in F. Note also the longer oligodendrocyte processes in the TNC null
cells (C) as compared with the WT cells (B). Cells were counterstained by DAPI
(blue). (F) More neurones develop from TNC null striatal neural stem cells.
The graph shows the increase in neurogenesis from TNC null cells in either
FGF2 or EGF. Results represent mean±s.e.m. of three independent
experiments; **P<0.01, using Student's t-test.
(G) The addition of exogenous TNC to neurosphere cultures from TNC-deficient
animals (–/–) reverses the increased neurogenesis to a level not
significantly different from that seen in heterozygous littermate controls
(+/–). Results represent mean±s.e.m. of three independent
experiments; *P<0.05, using Student's t-test.
Scale bars: 30 µm in B,C; 60 µm in D,E.
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