Figure 5.

Treatment with ERβ-specific agonists OSU-ERb-129 and LY500307 promotes anticancer effects in ERα+ breast cancer lines in vitro. (A) Colony formation. Colonies were stained with crystal violet and counted. The percentage of colonies present in each treatment is shown relative to DMSO vehicle-treated controls. Data are from three independent experiments and presented as mean ± SD; *p < 0.05, **p < 0.01, ***p < 0.001; n = 3. (B) Cell migration. Cell migration was determined using the wound-healing assay. The percentage of the filled area is calculated, normalized to DMSO-treated control, and presented as mean ± SD from three independent experiments; mean ± SD; *p < 0.05, **p < 0.01, ***p < 0.001; n = 3. (C) Enhanced cleavage of PARP-1, and activation of caspases 3 and 7 in ERα+ breast cancer cells upon treatment with ERβ agonists. Western blot analyses were performed using specific antibodies in whole-cell lysates prepared from OSU-ERb-12- and LY500307-treated cells as indicated. Similar results were obtained in different batches of cells treated with OSU-ERb-12 and LY500307. Numbers under the lanes are the quantitative representation of the intensity of the normalized bands. The signal in each band was quantified using Image Studio (LiCor) software. (D) Enhanced expression of FOXO1 and FOXO3a proteins in ERα+ breast cancer cells upon treatment with ERβ agonists. Western blot analyses were performed using specific antibodies in whole cell lysates prepared from cells treated for 7 days with OSU-ERb-12 or LY500307. Similar results were obtained with different batches of cells treated with OSU-ERb-12 or LY500307. Numbers under the lanes represent corresponding normalized band intensity of the respective proteins. Image Studio (LiCor) software was used to quantify the intensity of the protein bands.