Fig. 4. Recruitment of LFY and UFO to the AP3 promoter. (A) The AP3 genomic region. Three different regions of the AP3 promoter, DEE (distal early element), PEE (proximal early element) and INT (inter-region between DEE and PEE) (Hill et al., 1998) are illustrated. (B) (Left) Chromatin immunoprecipitation (ChIP) was performed with anti-FLAG antibody or mouse normal IgG serum from inflorescence tissue of plants of the indicated genotypes. Promoter regions from AP1 and AP3 were amplified using PCR as indicated. A region of the Mu transposon was used as a positive control for amplification. LFY specifically associates with both the DEE and PEE elements of the AP3 promoter, as well as with the AP1 promoter fragment. The ufo-2 mutation does not compromise the ability of LFY to bind to target sequences. (Right) DEE and PEE levels were normalized to Mu and the fold change of experimental IP over IgG control IP is indicated. The values are mean±s.e.m. from three PCR experiments. (C) (Left) ChIP was carried out using inflorescence tissues obtained from genotypes as indicated. Chromatin immunoprecipitations carried out using anti-Myc antibody or normal mouse IgG serum. UFO specifically associates with both AP3 promoter regions, DEE and PEE; however, the presence of the lfy-26 mutation abolished these interactions, indicating that LFY is required for UFO to associate with AP3 promoter sequences. (Right) Quantitation as in B.