Fig. 4

Deletion of the foxo1a gene leads to abnormal antioxidant systems in the zebrafish brain and ferroptosis occurs in the zebrafish brain.

A: ROS fluorescence images of the brains of control group and foxo1a^−/− group zebrafish larvae, scale bar = 100 μm

B: (a) The effect of antioxidant enzyme activity in the brain tissue of control group and foxo1a^−/− group zebrafish larvae, including SOD activity, CAT activity, GSH activity and MDA content. N = 6, t-test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. (b) IBR calculation radar chart of antioxidant enzyme activity in the brain tissue of control group and foxo1a^−/− group zebrafish larvae. N = 6, data are expressed as mean ± SD.

C: Iron ion fluorescence images of the brains of control group and foxo1a^−/− group zebrafish larvae, scale bar = 100 μm.

D: qRT-PCR detection of changes in oxidative stress-related mRNA expression (sod, cat, ho1, gpx4 and nrf2) in the brain tissue of control group and foxo1a^−/− group zebrafish larvae. N = 6, t-test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

E: qRT-PCR detection of changes in ferroptosis-related gene mRNA expression of hamp1, tfr1, dmt1, slc7a11 and nmdar in the brain tissue of control group and foxo1a^−/− group zebrafish larvae. N = 6, t-test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

F: (a) Flow cytometry detection of apoptosis in the brains of control group and foxo1a^−/− group zebrafish larvae. (b) The number and proportion of early apoptotic, late apoptotic, and necrotic cells in the brains of control group and foxo1a^−/− group zebrafish larvae, t-test, ∗p < 0.05, ∗∗p < 0.01.