Journal: The Journal of Biological Chemistry
Article Title: Ciprofloxacin enhances RSL3-induced ferroptosis by promoting mitochondrial Zn 2+ accumulation via the STING1–CAV2 pathway
doi: 10.1016/j.jbc.2025.110653
Figure Lengend Snippet: Ciprofloxacin (CFX) amplifies RSL3-driven ferroptosis. A, cell viability of PANC1, SW1990, and HeLa cells following treatment with different concentrations of CFX for 24 h. B, cell viability of PANC1, SW1990, and HeLa cells treated with RSL3 in the presence or absence of CFX (50 μg/ml) and liproxstatin-1 (Lip-1, 1 μM) for 24 h. C, representative Hoechst 33342 and propidium iodide (PI) staining images of PANC1, SW1990, and HeLa cells treated with RSL3 (1 μM) in the presence or absence of CFX (50 μg/ml) and Lip-1 (1 μM) for 24 h. Scale bar represents 200 μm. Quantification of PI-positive cells is shown. D, lipid reactive oxygen species (ROS) of PANC1 and SW1990 cells treated with RSL3 (1 μM) in the presence or absence of CFX (50 μg/ml) and ferrostatin-1 (Fer-1, 1 μM) for 12 h. E, malondialdehyde (MDA) of PANC1 and SW1990 cells treated with RSL3 (1 μM) in the presence or absence of CFX (50 μg/ml) for 12 h. F, fluorescence imaging of Phen Green SK (Fe 2+ indicator) in PANC1 and HeLa cells treated with RSL3 (1 μM) in the presence or absence of CFX (50 μg/ml) for 12 h. Scale bar represents 200 μm. The relative fluorescence intensity is shown. G, GSH level of PANC1 cells treated with RSL3 (1 μM) in the presence or absence of CFX (50 μg/ml) for 12 h. H, cell viability of PANC1 and HeLa cells treated with CFX (50 μg/ml) in the presence or absence of RSL3 (1 μM), staurosporine (STS, 1 μM), and TSZ (tumor necrosis factor-α, SM-164, and Z-VAD-FMK) for 24 h. Cell viability of PANC1 and HeLa cells treated with STS (1 μM) and TSZ (TNF-α, SM-164, and Z-VAD-FMK) in the presence or absence of the corresponding inhibitors (Z-VAD-FMK, 20 μM; necrosulfonamide/NSA, 2 μM) for 24 h. Data are presented as the mean ± SD from at least three independent experiments. p Values were calculated using two-way ANOVA ( B ) or one-way ANOVA ( C – H ). ns means no significance.
Article Snippet: Human tumor cell lines SW1990 (CRL-2172), PANC1 (CRL-1469), HeLa (CCL-2), and OVCAR-3 (HTB-161) were procured from the American Type Culture Collection and examined every 3 months to ensure they remained free of mycoplasma contamination.
Techniques: Staining, Fluorescence, Imaging