FIGURE 5.

ACTN4 associates with the pS2 promoter. A, association of ERα, ACTN4 and HDAC7 with the pS2 promoter. Chromatin prepared from MCF-7 cells treated with 10 nm E2 for the indicated time periods was subjected to chromatin immunoprecipitation assays as detailed under “Experimental Procedures,” with antibodies against ERα, HDAC7, and ACTN4. B, effects of overexpressing HDAC7 on ERE reporter activity. MCF-7 cells were transfected with reporter constructs harboring an ERE and β-gal along with increasing amounts of an HDAC7 expression plasmid. Luciferase reporter assays were performed as described in Fig. 2C. C, an alignment between the ACTN4 CaM-like domain and parts of calmodulin (CaM2). The CaM-like domains of all four ACTNs are similar to each other. D, ACTN4 (Δ831–869) loses its ability to potentiate ERE reporter activity. The reporter assays were (panels B and D) were performed as described in Fig. 2C.