Figure 7.

ML385 reverses the neuroprotective effects and resistance to ferroptosis in mice with CISD2 overexpression following OGD/R injury.

(A, B) The effect of CISD2 overexpression on HT22 cell survival rates following OGD/R injury, evaluated via Calcein-AM/PI. The data were normalized with respect to the control group. Compared with the OGD/R + OE group, the cell survival rates were significantly reduced in the OGD/R + OE + ML385 group. Green: living cells, red: dead cells. (C–F) Representative western blots and quantitative evaluations of the effect of ML385 on the expression of TFR1, xCT, and GPX4 following OGD/R injury. (G) The effect of ML385 on the fluorescence of Fe²⁺ following OGD/R injury. The levels of Fe²⁺ were significantly increased in the OGD/R + OE + ML385 group when compared with those in the OGD/R + OE group. Scale bars: 100 µm (A), 10 µm (G). (H, I) The effect of ML385 on the lipid ROS following OGD/R injury, detected via flow cytometry. Data are represented as the mean ± SEM (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (one-way analysis of variance followed by Tukey’s post hoc test). CISD2: CDGSH iron sulfur domain 2; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GPX4: glutathione peroxidase 4; ML385: inhibitor of Nrf2; NC: negative control; OGD/R: oxygen-glucose deprivation/reoxygenation; OE: overexpression; ROS: reactive oxygen species; TFR1: transferrin receptor 1; xCT: a cystine/glutamate antiporter.