Fig. 2

Cytotoxicity, copper ion dynamics, and molecular mechanism analysis of copper ions, elesclomol, and their combination in epithelial and fibroblast cells. A CCK-8 assay showing cell viability in three groups: copper ion (Cu) alone, elesclomol alone, and Cu&elesclomol. Cells were treated with increasing concentrations of Cu and/or elesclomol (0, 1, 10, 50, 100, 200, 500, 1000, 5000, 10,000 nM) for 12 h. B Time-dependent cytotoxicity of the Cu&elesclomol group at concentrations of 100 nM, 200 nM, and 500 nM after 0, 12, 24, and 36 h, assessed via CCK-8 assay. C Effects of inhibitors on Cu&elesclomol-induced cytotoxicity. Treatments include Cu&elesclomol (1:1, 500 nM) alone or in combination with Z-VAD–FMK (30 μM), Necrostatin-1 (20 μM), or TTM (20 μM) for 0, 12, and 24 h, evaluated via CCK-8 assay. D Intracellular copper content in cells in four groups: CT, copper ion alone, elesclomol alone, and Cu&elesclomol. E Western blot (WB) analysis of Olig-DLAT expression in epithelial cells (Beas-2b) from the control (CT) group and Cu&elesclomol-treated group. F WB analysis of epithelial–mesenchymal transition (EMT) markers E-cadherin and Vimentin in epithelial cells (Beas-2b) from the CT group and Cu&elesclomol-treated group, with quantitative analysis. G WB analysis of fibrotic markers α-SMA and Col1 A1 in fibroblast cells (MRC-5) from the CT group and Cu&elesclomol-treated group, with quantitative analysis. Data are presented as mean ± SD.*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (two-way ANOVA or t tests) in this figure