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Despite these pH-dependent artifacts, the literature does not clearly define an optimal pH working range for the assay

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Moreover, increased intracellular copper levels activate multiple signaling pathways, such as mitogen-activated protein kinase/extracellular signal-regulated kinase (MEK/ERK) and B-rapidly accelerated fibrosarcoma (BRAF),which stimulate cell proliferation, promote angiogenesis via vascular endothelial growth factor (VEGF) activation (Pan et al., 2002), and enhance tumor invasion and metastasis by inducing epithelial-mesenchymal transition (EMT) (Zhang et al., 2025)

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