Fig. 4.

Effect of RANKL, denosumab, and trastuzumab in NF-κB signaling pathway in BC cells. a–d Cells were starved and treated with RANKL, denosumab, and/or trastuzumab. Western blot analysis from cell lysates of SKBR3, MCF7, MDA-MB-453, and BT-474 using phospho-IKKα, IKKα, phospho-IκBα, IκBα, phospho-NF-κB p65, and NF-κB p65 antibodies. Actin was used as a protein loading control. Representative Western blots of each cell line are presented. e–h The histograms represent densitometry results of the phospho-immunoblots normalized by total protein levels. Data were analyzed by one-way ANOVA and represent mean ± SD. Asterisks indicate *p < 0.05, **p < 0.01, and ***p < 0.001