Fig. 7: Bufalin overcomes Tamoxifen resistance in vitro.

a Representative IHC staining for ERα in Tamoxifen treatment relapsed simple (Scale bar 20μm). b Tamoxifen-resistant cells LCC2 were treated with a serious of Bufalin, the expression of ERα was measured by western blot. c Tamoxifen-resistant cells LCC2 were treated with Bufalin, and cell viability was determined using the CCK-8 assay, the data are presented as mean ± SD, n = 3 independent experiments. One-way ANOVA was used for statistical analysis, P < 0.05 was considered statistically significant. d The colony formation assay was used to measure LCC2 cell proliferation after treatment by Bufalin, the data are presented as mean ± SD, n = 3 independent experiments. One-way ANOVA was used for statistical analysis, P < 0.05 was considered to be statistically significant. e The EdU assay was used to measure LCC2 cell proliferation following treatment with Bufalin (Scale bar 100μm), the data are presented as mean ± SD, n = 3 independent experiments. One-way ANOVA was used for statistical analysis, P < 0.05 was considered statistically significant. f LCC2 cells were treated with 50 or 100 nM Bufalin for 48 h, and the expression of PARP and Bcl-2 was measured by western blot, the samples derive from the same experiment and that gels/blots were processed in parallel. g LCC2 cells were treated with 50 or 100 nM Bufalin for 48 h, and apoptosis was examined by measuring Annexin V staining, the data are presented as mean ± SD, n = 3 independent experiments. One-way ANOVA was used for statistical analysis, P < 0.05 was considered statistically significant. h The colony formation assay was used to measure LCC2 cell proliferation after treatment with Bufalin or Fulvestrant, the data are presented as mean ± SD, n = 3 independent experiments. One-way ANOVA was used for statistical analysis, P < 0.05 was considered statistically significant. i Tamoxifen-resistant cells LCC2 were treated with Bufalin and Fulvestrant, and cell viability was determined using the CCK-8 assay, the data are presented as mean ± SD, n = 3 independent experiments. Two-way ANOVA was used for statistical analysis, P < 0.05 was considered statistically significant.