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Fig. 4. Sox9 directly activates Snail2 promoter.
(A) Activation of Snail2 promoter by Sox9 in neural
plate explants. D1.2-Luciferase reporter gene and other
`crest-specific' transcription factors were electroporated into neural plates,
and cultured for 20 hours without BMP4. Only Sox9 can activate the
reporter. Results are shown as fold-induction compared with the result
obtained from explants, transfected with D1.2-Luciferase and empty
pyDF30 plasmid. (B) Sox9 binds to the flanking sequence of E-box2. EMSA
was performed to determine the binding site of Sox9 on Snail2
promoter. The sequence and the position of the probe and competitors are shown
below. E-box2 is indicated by an open box. The Sox9 binding to the E-box probe
is interfered by the addition of competitor 1, 3 and 5, but not by competitor
2 or 4. (C) The flanking sequence of E-box2 is required for the
Snail2 promoter activation by Sox9. When the C-rich sequence next to
the E-box2 in the D0.1 reporter is mutated (Em3), activation level of the
promoter by Sox9 (+) is significantly decreased, compared with the wild-type
D0.1. (D) Sox9 and Snail2 synergistically activate
D0.1-Luciferase promoter. D0.1-Luciferase was co-transfected
with Sox9 or Snail2 (or both) into NIH3T3 cells. Results are
shown as a fold-induction compared with the result obtained from cells
transfected with D0.1-Luciferase and an empty pyDF30.
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