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. 1999 Jun 8;96(12):6683–6688. doi: 10.1073/pnas.96.12.6683

Figure 1.

Figure 1

ZEB and zfh-1 bind to CtBP-1 and CtBP-2. (A) ZEB and zfh-1 contain CtBP binding sites. Drosophila zfh-1 and ZEB from various vertebrate species (h: human; ck: chicken; hms: hamster; ms: mouse and rat) contain a PLDLS CtBP binding sequence in their repressor domain. In addition, vertebrates contain two additional PLDLS-like sequences, which vary slightly from species to species. (B) Flag-tagged constructs for snail, the repressor domains of ZEB and zfh-1 (the region between both zinc finger domains) as well as the DNA binding domains (C-terminal zinc fingers) of ZEB (DB-ZEB) and zfh-1 (DB-zfh-1) were cotransfected in C33a cells with myc-tagged CtBP-1. After 48 hr, cells were lysed and after immunoprecipitation with Flag antibody, binding to CtBP-1 was detected by Western blot using 9E10 anti-myc mAb as described in Materials and Methods. Ten percent of the lysate was run without immunoprecipitation as input control. Blots then were stripped and incubated with anti-Flag antibody to detect levels of snail, ZEB and zfh-1. (C) As in B but using myc-tagged CtBP-2.