Fig. 3: IMT1 disrupts mitochondrial functions in OS cells.

The primary OS cells, pOS-1, were treated with IMT1 (1 μM) for 24 h, mitochondrial depolarization was analyzed by JC-1 staining (A); ROS production was tested via CellROX and DCF-DA dye assays (B, C), with lipid peroxidation examined via BODIPY staining (D); The ssDNA contents were measured via ELISA assay (E); The mitochondrial complex I activity (F) and ATP contents (G) were also measured. pOS-1 cells were pretreated for 1 h with NAC (500 μM), ATP (1 mM) or PBS, followed by IMT1 (1 μM) treatment for indicated time periods, cell viability, death and apoptosis were tested by CCK-8 (H), Trypan blue staining (I) and nuclear TUNEL staining (J), assays, respectively. Other primary OS cells (pOS-2 and pOS-3) or the immortalized MG63 cells were treated with IMT1 (1 μM) for 24 h, mitochondrial depolarization (JC-1 dye assay, K), ROS production (CellROX dye assay, L) and ATP contents (M) were tested similarly. “Veh” designates the vehicle control group. *P < 0.05 indicates a significant difference compared to the “Veh” treatment. ^# P < 0.05 versus “PBS” group (H–J). The data is presented as mean ± SD. The experiments presented in this figure were replicated five times (n = 5, with five biological replicates), yielding consistent and similar results. The scale bar is set at 100 μm.