Research Paper Volume 15, Issue 24 pp 14900—14914

Figure 5. Acetylshikonin induced necroptotic lipid peroxidation in NSCLC cells. (A) Transmission electron microscopy analysis showing impaired membrane integrity (blue arrow) after treating H1299 cells with acetylshikonin for 6 h. The red arrow indicates mitochondria, and the white arrow indicates lysosomes. 5,000×: Scale bar = 2 μm. 20,000×: Scale bar = 0.5 μm. (B–D) Fluorescence microscope images and flow cytometry results indicating lipid peroxidation in H1299 and A549 cells incubated with acetylshikonin (0.5–10 μM) and BODIPY™ 581/591 C11 for 30 min (n = 4). Scale bar = 200 μm. (E, F) Western blot analysis showing GPX4 protein expression in H1299 and A549 cells following treatment with acetylshikonin (0.5–10 μM) for 24 h (n = 4). Untreated cells were used as controls. Results are shown as means ± SD. ^*p < 0.05 compared to untreated control.