Analysis of FOXO binding sites by chromatin immunoprecipitation. T98G cells were transfected with an empty pcDNA3 control or Flag-FOXO3a-AAA expression vector for 24 hours. Chromatin fragments were immunoprecipitated with anti-Flag antibody and quantified by real-time PCR (see Additional file 4 for primers). Data are presented as percentage of input and are the means of 3 independent transfections ± S.E β-globin is a negative control. Immunoblots indicated that the expression of FOXO3a-AAA in transfected cells was 3–5 fold greater than the endogenous protein.
Terragni et al. BMC Cell Biology 2008 9:6 doi:10.1186/1471-2121-9-6