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. 2022 Jun 6;25(7):104533. doi: 10.1016/j.isci.2022.104533

Figure 2.

Figure 2

Myriocin inhibits oxidative damage induced by erastin or glutamate treatment

HT22 cells were pre-treated with or without myriocin (0.5 μM) or DHS (1 μM) for 36 h before incubating with or without erastin (1 μM) or glutamate (15 mM) for 24 h.

(A) Intracellular glutathione levels of cells treated as indicated.

(B) Lipid peroxidation of cells treated as indicated and monitored by flow cytometry after labeling with C11-BODIPY 581/591, fluorescence intensity was measured on the FITC channel (left two panels: distributions of fluorescence, right panel: relative quantification of fluorescence enhancement which represents the extent of probe oxidation).

(C) Intracellular ROS production of cells treated as indicated and visualized by using the fluorescent probe DCFH-DA, scale bar: 50 μm. Bar graph shows the quantification of fluorescence intensity relative to the relevant untreated control.

(D and E) Relative level of labile iron pool (LIP) in cells treated as indicated and monitored by the calcein fluorescence quenching method. For the above, error bars represent the mean ± SD (n = 3, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001).