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Fig. 6. The Pho consensus site is required for Pho binding in vivo. ChIP
analysis of Pho binding (as in Fig.
5) to transgenes carrying PRE300 (GFP-RR,
Fig. 2), either with a normal
(A-C) or a mutated (D-F) Pho site, in 0-15 hours AED embryos
(eggs were collected for 15 hours at temperature; embryos spanned stages 1 to
late 16). The signal from the transgene (`wt t'gene' or `mut t'gen', using a
transgene-specific primer set) is compared in each case with that from the
endogenous eve 3' PRE (`endo eve', primer set 13 of
Fig. 5), as well as to a gene
(actin) that Pho is not expected to bind. Note that the Pho signal is reduced
when the Pho site is mutated (transgene signal in D versus that in A),
relative both to the endogenous eve signal and to the (presumably
background) signal from the actin gene.
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