Fig. 4.

Sf1 SUMOylation and Dax1 modify Sf1 activity on the FAdE enhancer in vitro. (A) Ad4 (Sf1) binding sites in the mouse FAdE region (Zubair et al., 2002). (B) Binding capacity of Sf1 on the different Ad4 sites in the FAdE region. ChIP assays were performed on Y1 cell lines using anti-Sf1 antibodies. Immunoprecipitates were analyzed by quantitative PCR using primers designed for each individual site. The data were normalized to values obtained for 1% input controls, and the results are presented as percentage of input. Data are mean±s.e.m. (C) SUMOylation and Dax regulate Sf1 activity on the FAdE enhancer. HEK293T cells were plated at 10⁵ cells/well in 24-well plates 24 h before transfection and were transfected in triplicate with FAdE Luc (100 ng/well), with Sf1 as indicated, and with or without Dax1 (50 ng/well). Luciferase assays were carried out and the data were normalized to renilla level and shown as fold change over control vectors. n=6. *P<0.05. Error bars indicate s.e.m.