Figure 8.

(a) Western blot analysis of Jun and Fos proteins carried out using nuclear extracts from untreated and E₂-treated RAW 264.7 cells stimulated with IL-1 and TNF for 1 hour and polyclonal antibodies against all members of the Jun and Fos families of nuclear proteins. Representative data from 1 of 3 replicate experiments. (b) Western blot analysis conducted using antibodies against specific Jun proteins demonstrated that E₂ downregulates c-Jun and JunD levels, but not JunB levels. Representative data from 1 of 3 replicate experiments. (c) E₂ decreases c-Jun gene expression in IL-1– and TNF-stimulated RAW 264.7 cells. Control- and E₂-treated RAW 264.7 cells stimulated with IL-1 and TNF were transiently transfected with a c-Jun/luciferase construct containing the full-length c-Jun promoter. Luciferase activity was normalized to β-galactosidase activity to correct for variability in transfection efficiency and expressed as normalized luciferase activity. Mean ± SEM of 3 experiments. *P < 0.05 vs. untreated cells.