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Cell viability levels of SK‐BR‐3 and MCF‐7 determined by MTT assay. (A, B) Effects of single (A) and in combination (B) therapeutic agents on cell viability in HER2 + (SK‐BR‐3) and HER2 − (MCF‐7) breast cancer (BC) cell lines. The data is expressed as mean ± SD of three independent experiments ( N = 3). Statistical significance was determined by ANOVA followed by Dunnett's T3 post hoc test, where * p ‐value < 0.05; ** p ‐value < 0.01; *** p ‐value < 0.001 in comparison with the control group. (C) Experiments at the level of 3D cell lines, when incubated with D+T+P+N+I (Denosumab+Trastuzumab+Pertuzumab+Nivolumab+Ipilimumab) in HER2 + BC, resulted in a clinically significant reduction in cell viability, possibly through immunomodulation of the tumour microenvironment (TME), characterised by a marked increase in tumour‐infiltrating lymphocytes (TILs) infiltration and a possibly concurrent decrease in pro‐tumorigenic neutrophils (created using BioRender ( https://biorender.com/ )). ANOVA, analysis of variance; MTT, 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide; SD, standard deviation.

Journal: Journal of Cellular and Molecular Medicine

Article Title: The Role of HER2 and RANK in Breast Cancer and New Therapeutic Approaches With Denosumab, Anti‐HER2 Antibodies and Immunotherapy

doi: 10.1111/jcmm.71071

Figure Lengend Snippet: Cell viability levels of SK‐BR‐3 and MCF‐7 determined by MTT assay. (A, B) Effects of single (A) and in combination (B) therapeutic agents on cell viability in HER2 + (SK‐BR‐3) and HER2 − (MCF‐7) breast cancer (BC) cell lines. The data is expressed as mean ± SD of three independent experiments ( N = 3). Statistical significance was determined by ANOVA followed by Dunnett's T3 post hoc test, where * p ‐value < 0.05; ** p ‐value < 0.01; *** p ‐value < 0.001 in comparison with the control group. (C) Experiments at the level of 3D cell lines, when incubated with D+T+P+N+I (Denosumab+Trastuzumab+Pertuzumab+Nivolumab+Ipilimumab) in HER2 + BC, resulted in a clinically significant reduction in cell viability, possibly through immunomodulation of the tumour microenvironment (TME), characterised by a marked increase in tumour‐infiltrating lymphocytes (TILs) infiltration and a possibly concurrent decrease in pro‐tumorigenic neutrophils (created using BioRender ( https://biorender.com/ )). ANOVA, analysis of variance; MTT, 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide; SD, standard deviation.

Article Snippet: Following treatment, the reagent 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) (30006, Biotium Inc., Landing Parkway, Fremont, CA, USA) was added to each well at a final concentration of 0.5 mg/mL.

Techniques: MTT Assay, Comparison, Control, Incubation, Standard Deviation