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. 2000 Jan 15;28(2):e4. doi: 10.1093/nar/28.2.e4

Figure 3.

Figure 3

Zeste is crosslinked most strongly to a 297 bp region of the Ubx promoter. (A) Diagram of the restriction fragments produced by various digests of the Ubx proximal promoter. The positions of EcoRI (R), EheI (E), MluI (M) and StuI (S) restriction sites are marked, and the RNA start site at +1 is indicated by an arrow. The restriction fragments to which binding has been examined are a 2.8 kb EcoRI–StuI fragment that spans nucleotides –3.54 kb to –677 bp (a), a 1 kb EcoRI–StuI fragment from –677 to +365 bp (b), a 3.4 kb EcoRI–EheI fragment from –3.54 kb to –177 bp (c), a 540 bp EcoRI–EheI fragment from –177 to +365 bp (d), a 1.8 kb EheI–EheI fragment from –177 bp to +1.63 kb (e) and a 1.5 kb MluI–EheI fragment from +120 bp to +1.63 kb (f). Ovals denote the positions of high affinity Zeste binding sites. (B) Results of formaldehyde crosslinking experiments examining binding to the DNA fragments shown in (A). Vertical columns show blots of immunoprecipitations of crosslinked chromatin from wild-type 0–12 h embryos (a–f). The top row contains DNA fragments immunoprecipitated with anti-Zeste antibodies, the middle row contains the result obtained with non-specific antibody and the bottom row contains 1% of the input DNA. The Southern blots examining binding to fragments a–d were probed with the same 3.5 kb fragment of the Ubx proximal promoter as used in Figure 2A. The blots examining binding to fragments e and f were probed with an adjacent EcoRI DNA fragment that lies between nucleotides +365 bp and +3.68 kb.