Fig. 4: Knockdown of ZIP8 enhances ferroptosis in ESCC cells.

A Functional schematic illustrating the role of ZIP8 in the ferroptosis pathway. B Sensitivity of ESCC cells to the ferroptosis activator Erastin, assessed by CCK-8 assay following treatment with increasing concentrations of Erastin (0 µM, 0.01 µM, 0.1 µM, 1 µM, 10 µM). Data represent mean ± SD of n = 3 independent experiments. C Lipid peroxidation assay to evaluate oxidative damage, with malondialdehyde (MDA) concentration measured using an MDA assay kit. D Western blot analysis of FTL and FTH1 expression in ESCC cells following ZIP8 knockdown. E, F Detection of lipid peroxidation in ZIP8 knockdown cells using the C11-BODIPY fluorescent probe. Scale bar, 20 µm. G, H Assessment of lipid peroxidation in KYSE30 and KYSE510 cells using the BD FACSAria II flow cytometer with the BODIPY™ 581/591 C11 probe. I, J Measurement of intracellular ferrous iron (Fe²⁺) concentration in ZIP8 knockdown cells using the FerroOrange fluorescent probe. Scale bar, 20 µm. Student’s unpaired t-test was employed for statistical analysis in (B, C, F, H, J). *p < 0.05, **p < 0.01, ***p < 0.001 (n = 3, independent experiments). Data are represented as mean ± SD.