Fig. 6
From: Combining RANK/RANKL and ERBB-2 targeting as a novel strategy in ERBB-2-positive breast carcinomas
Effect of pertuzumab addition to denosumab plus trastuzumab in ERBB2-positive BC cells. Cells were starved and treated as indicated. a Representative images are shown using confocal microscopy. Red dots represent the positive signal of RANK/ERBB2 heterodimers in SKBR3 and BT-474 cells using DUOLINK in situ proximity ligation assay (PLA). Scale bars represent 20 μm. b Dot plots represent the total signal per cell as analyzed by Duolink Image Tool Software. c Western blot analysis from cell lysates of SKBR3 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. d The histograms represent densitometry results of the phospho-immunoblots normalized by total protein levels. e XTT proliferation assay for SKBR3 and BT-474 cells, after 24 and 72 h of treatment. Results are expressed in the histogram as growth inhibition, normalized to the control group. f Quantification of migration-wound healing assay for SKBR3 and BT-474 cells analyzed at 24 and 48 h. The histogram shows percent wound recovery at 24 and 48 h in relevance to 0 h. Data in b–f were analyzed by one-way ANOVA and represent mean ± SD. Asterisks indicate *p < 0.05, **p < 0.01, and ***p < 0.001