Fig. 3. Analysis of Estrogen-Dependent Effects after Perturbation of Bip Expression in the Primary Culture of Uterine Stromal Cells.

A, Determination of endogenous ERα-mediated transactivation of luciferase activity. Cell extracts were analyzed by luciferase activity as described in Materials and Methods. Data (as percent) are represented by mean ± sem of relative luciferase activity from four independent experiments. Data were analyzed using Student’s paired t test (P < 0.01). B, Analysis of cellular growth. Primary cultured cells were subjected to infection with 1 pfu/cell of adenoviruses (rAdBipAs or rAdGFP) for 12 h, followed by the treatment of E₂ (10 nm) for indicated times. Noninfected cells were also analyzed in parallel with or without addition of estrogen. Cellular proliferation was assayed as described in Materials and Methods. The asterisks (**) indicate statistically different (P < 0.01; ANOVA followed by Newman-Keul’s multiple-range test) as compared with corresponding control group on particular days. C, Analysis of cellular viability. Cells on d 3 as examined in panel B were visualized by fluorescence microscopy after staining with 4’,6-diamidino-2-phenylindole (DAPI) and propidium iodide (PI). Viable cells exclude PI, whereas dead cells stain with PI (red) and DAPI (blue).