Figure 1. Norgestrel increases LIF in 661W cells. A: 661W cells were screened for relative changes in mRNA expression of the neurotrophic factors leukemia inhibitory factor (Lif), epidermal growth factor (Egf), hepatocyte growth factor (Hgf), and nerve growth factor (Ngf), and basic fibroblast growth factor (bFGF) following 20 µM norgestrel treatment for the times shown or vehicle control (ctrl) for 6 h. B: Relative expression of Lif in 661W cells treated with 20 µM norgestrel or vehicle control for the times indicated. For A and B, relative expression was analyzed with real-time (RT) PCR, with the fold change compared to the geometric mean of the three endogenous reference genes. C: Western blot analyses confirm an increase in LIF at the protein level following treatment with 20 µM norgestrel or vehicle control. Densitometry was calculated from three individual experiments and graphed as a percentage of the control. A representative blot is shown. Tubulin acted as the loading control. D: Immunofluorescent staining also shows an increase in LIF immunoreactivity (green) in 661W cells following norgestrel treatment for the times indicated or vehicle control for 6 h. Nuclei were stained with Hoechst dye. Scale bar represents 50 µm. Error bars denote ± standard error of the mean (SEM) from three independent experiments. **p<0.01, *** p<0.001, ****p<0.0001 (ANOVA).