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. Author manuscript; available in PMC: 2018 Jun 1.
Published in final edited form as: Mol Cancer Ther. 2017 Mar 3;16(6):1080–1091. doi: 10.1158/1535-7163.MCT-16-0626

Figure 1. G-1 inhibits the proliferation of breast cancer cells independent of the expression status of GPER and classic estrogen receptors.

Figure 1

A) G-1 inhibits the proliferation of breast cancer cells in a concentration-dependent manner. Cells were treated with increasing concentrations of G-1 for 48 h. Cell numbers were counted using an automatic cell counter and presented as percentages of that of control group (0.1% DMSO treated). Each bar represents mean ± SEM of four repeats. Bars with different letters are significantly different from each other (P < 0.05). B) G-1 inhibits the proliferation of breast cancer cells in a time-dependent manner. Cells were treated with 2 μM G-1 for different time. Cell numbers were presented as percentages of that of control group (0 h group). Each bar represents mean ± SEM of four repeats. Bars with different letters are significant different from each other (P < 0.05). C) Representative Images showing colony formation of breast cancer cells in the presence of different concentrations of G-1 for 7 days. Quantitative data show the colony numbers. Colonies were stained with MTT. Scale bar: 200 μm. CTRL: 0.1% DMSO treatment. ***: P < 0.001 compared to control. D) Representative images showing the nude mice bearing breast cancer xenograft treated with or without G-1 for 2 weeks (left panel), and the harvested tumors (right panel). Red circles indicate xenograft location and size. CTRL: sesame oil control treatment every other day for 2 weeks; G-1: 5 mg/kg G-1 treatment every other day for 2 weeks. E) Tumor xenograft growth curves in athymic nude mice treated with or without G-1. The xenograft volume was measured weekly. Student’s T test was used to compare the tumor volumes between control and G-1 treated groups. Each point represents mean ± SEM (n = 6). *: P < 0.05. F) The weight of tumor xenografts from nude mice treated with or without G-1 for 2 weeks. Each bar represents mean ± SEM (n = 6). **: P < 0.01 compared to control. G) Representative images showing expression of Ki67 in control and G-1 treated tumor xenografts determined by fluorescent immunohistochemistry. Ki67 was labeled in green; Actin was stained with rhodamine-phalloidin (red); Nuclei were stained with DAPI (blue). Scale bar: 20 μm. The number of Ki67 positive cells and total cells in control and G-1 treated groups were also counted under a fluorescent microscope and the ratio of Ki67 positive cells was calculated. Each bar represents mean ± SEM (n = 6), ***: P < 0.001. H) The body weight of athymic nude mice with or without G-1 treatment. Each bar represents mean ± SEM (n = 6). Student’s T test was used to do statistical analysis. No statistical significance on the body weight was observed between control and G-1 treated groups.