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.