Fig. 2

rLCN2 induced ferroptosis in 661 W photoreceptor cells. A Cell viability of 661 W cells incubated with serial concentrations of rLCN2 (0, 0.1, 1, and 10 μg/mL) for 24 h. B‒D Western blotting and quantitative analysis of SLC7A11 and GPX4 protein expression levels in 661 W cells exposed to 1 μg/mL rLCN2 for 24 h. The protein expression levels of SLC7A11 and GPX4 were normalized to those of β-actin and are presented as fold changes. E TEM images of mitochondria (arrows) in 661 W cells treated with 1 μg/mL rLCN2 for 24 h. Scale bars = 1 μm. F Intracellular Fe²⁺ levels, measured using a colorimetric iron assay kit, in 661 W cells treated with 1 μg/mL rLCN2 for 24 h. G Intracellular ROS levels, measured using DCFH-DA (green), in 661 W cells treated with 1 μg/mL rLCN2 for 24 h. Nuclei were stained blue with Hoechst 33342 dye solution. Scale bars = 50 μm. H Quantification of ROS levels as the proportion of green cells (%). I MDA levels in 661 W cells incubated with 1 μg/mL rLCN2 for 24 h. J GSH levels in 661 W cells after incubation with 1 μg/mL rLCN2 for 24 h. GSH levels are shown as a percentage of the levels in control cells. Cells treated with vehicle (PBS) alone served as the control group. n = 3 per group. *P < 0.05, **P < 0.01. One-way ANOVA followed by Tukey’s post hoc test for A-D, Student's t‐test for F, H-J