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Fig. 7. SRF and GATA4 can simultaneously bind XMLC2 promoter in vitro, and can
synergistically activate an XMLC2-luciferase reporter in animal pole explants.
(A) The CArG-like site in 122/85 promoter fragment binds SRF
only weakly (complex labelled S); GATA4 protein binds efficiently to the
GATA#2 site present in the same probe (labelled G); simultaneous binding of
SRF and GATA4 can be detected (arrow). (B) Overexpression of SRF activates an
XMLC2(123/41) cyt.actin-luciferase reporter only weakly in
animal pole explants, while ectopic GATA4 gives strong activation.
Co-expression of both factors leads to synergistic activation. Animal poles
were excised at stage 9 and collected after three hours of incubation at
23°C. Firefly luciferase activity was normalised using coinjected
tk-Renilla luciferase as a control and was set at 1 for
XMLC2(123/41) cyt.actin-luciferase in the absence of SRF or
GATA4. Values shown represent a single experiment; a second experiment gave
similar results.