Supporting information for Loy et al. (2003) Proc. Natl. Acad. Sci. USA, 10.1073/pnas.0736237100
Fig. 7.
FLNa-mediated transcription repression is selective. Increasing amounts of FLNa were coexpressed with each steroid receptor in the presence of the appropriate reporter construct. The naturally occurring mouse mammary tumor virus-luciferase (MMTV-Luc) reporter gene was used to measure the transactivation activity of the progesterone (PR) and glucocorticoid receptors (GR), whereas a reporter regulated by a multimeric estrogen response element (ERE) promoter was used for the estrogen receptor. HeLa cells were transfected with increasing doses of pCMV-FLNa (0, 20, or 40 ng) together with 100 ng of MMTV-Luc and 50 ng of PR (A) or GR (B). Similarly, cells were transfected except that 50 ng of ERα (C) or ERβ (D) and 100 ng of ERE-Luc were used. Five hours after transfection, cells were treated with vehicle or with their appropriate ligand at 100 nM progesterone, 1 nM hydrocortisol, or 1 nM 17 β-estradiol. Activity of the steroid receptors was accessed by measuring the luciferase activity (RLU).