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Fig. 7. Activation of myelin gene expression by Sox8, and comparison of
ß-galactosidase activities in Sox8+/lacZ and
Sox10+/lacZ spinal cords. (A) N2A cells were transfected
with reporter plasmids in which the luciferase gene was under control of a
long (positions -656 to +31) or a short (positions -256 to +31) version of the
rat Mbp promoter. Expression plasmids for Sox10 and Sox8 were
co-transfected as indicated below the bars. Luciferase activities in extracts
from transfected cells were determined in three independent experiments each
performed in duplicates. Data are presented as fold inductions ±s.e.m.
with the activity of the promoter in the absence of co-transfected Sox protein
(-) arbitrarily set to 1. (B) RT-PCR analysis on cDNA obtained from N2A cells
inducibly expressing Sox8. - Doxy, no doxycycline (Sox8 absent); + Doxy,
doxycycline added (Sox8 present). Transcript levels of Sox8, Plp, Mbp
and ß-actin were compared semi-quantitatively using increasing numbers
(n, n+3, n+6) of amplification cycles. -, no cDNA added. (C) The amount of
ß-galactosidase present per µg extract from
Sox8+/lacZ (white bars) and
Sox10+/lacZ (black bars) spinal cords was determined at
14.5 dpc, 16.5 dpc, 18.5 dpc and in adult mice as indicated below the
bars.
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