Figure 6
Temporal changes in uterine cytokine mRNA and protein in response to E 2 . Uterine mRNA analysis via TaqMan® real-time PCR (panel A) and protein analysis via Western blot (panel B) are shown for TNF-α and MCP-1 from uteri of OVX rats 0-72 h after treatment with saline (0 h control) or E2 (40 μg/kg). Messenger RNA values (panel A) represent mean ± SE, n = 5-6 per group; one way ANOVA, *P < 0.05 compared to 0 h time point. Western blots images shown (panel B) are representative of a minimum of 3 blots using 3 separate pools of total protein extracted from different animals in 2 separate experiments. Intensities of protein bands were quantitated using AlphaEase FC software (Alpha Innotech) as described in the Methods and values for each time point expressed as percent of saline control (100%). Quantitative data (panel B) reflect average band intensity values ± SE measured from ≥ 3 blots with total protein extracted from different animals in 2 separate experiments; one way ANOVA, *P < 0.05 compared to 0 h time point. IDV = integrated density value. Western blot analysis of β-actin as a control for gel loading is shown for the protein samples used to assess TNF-α and MCP-1 protein for the blots above.