Fig. 2. Uterine regulation of gene specific mRNAs by xenoestrogenic compounds and E₂. Dose response studies. Adult ovariectomized wild-type mice were injected (sc) with (A), kepone (7.5, 15 or 30 mg/kg); (B), o,p’-DDT (7.5, 15 or 30 mg/kg); (C) methoxychlor (7.5, 15 or 30 mg/kg) or (D), E₂ (4 μg/mouse) (as a positive control) for 24 h. Mice injected with oil and killed after 24 h was served as vehicle control.

Total RNA (6 μg) was separated by formaldehyde-agarose gel electrophoresis, transferred and UV cross-linked to nylon membrane, and hybridized as described in the Materials and Methods. Hybridization was performed using 32P-labeled cRNA probes sequentially to Bip, SFRP-2 (secreted frizzled related protein 2) and rpL7 (ribosomal protein L7). Representative autoradiograms are shown with exposures at 5 h for Bip, 15 h for SFRP-2 and 3 h for rpL7. Temporal studies. Uterine total RNA (6 μg in each lane) was analyzed at indicated times, using kepone at 7.5 mg/kg (E) or 30 mg/kg (F) or E₂ (4 μg/mouse) (G). Hybridization was performed using ³²P-labeled cRNA probes sequentially to Bip, and rpL7. Autoradiographic exposure times were similar as indicated above. In general, 3-5 mice were used for each group analysis. These experiments (A-G) were repeated three times with independent RNA samples. In the bar plot, the abundance of mRNAs for each gene expression was quantitated by analysis of band intensities using densitometric scanning and was corrected against rpL7. *Values are statistically different (p<0.05, Student’s t test) from the oil treated group.