Figure 3.

ZEB1 binds with higher affinity to the murine Lhb than human LHB promoter. (A) Nuclear extract (NE) from HEK293T cells transfected with pcDNA3.0 or ZEB1-HA-FLAG was incubated with a biotinylated probe corresponding to −38/−15 of the murine Lhb promoter. Where indicated, NE was pre-incubated with control (IgG) or α-HA antibody. (B) NE was pre-incubated with unlabeled competitor probes corresponding to the indicated regions of the murine Lhb or human LHB promoter. (C) Quantification of EMSA assays in B. (D) NE was pre-incubated with unlabeled competitor probes corresponding to the indicated wild-type (WT) or Z-box or E-Box mutant (MUT) murine Lhb probes. (E) Quantification of EMSA assays in D. For B and D, increasing concentrations of unlabeled competitor probes are denoted by the gradient-filled slopes; 10X, 50X, and 250X molar excess. One representative experiment of 3 is shown. Data are the mean of 3 independent experiments. White circles represent experimental replicates. Data were analyzed by two-way ANOVA followed by Holm–Sidak multiple comparisons test. Bars with different letters differ significantly.