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. 2023 Oct 11;14(10):668. doi: 10.1038/s41419-023-06182-4

Fig. 4. IL-33/ST2 inhibited ferroptosis by regulating SLC7A11 expression in eESCs.

Fig. 4

A Pearson’s test was used to analyze the relationship between the levels of IL-33 and SLC7A11 mRNA in EC tissues (n = 8). B EESCs were treated with specified concentrations of GSH (0.5, 1.0, 1.5, 2.0, 2.5, and 3.0 mM) for 2 or 24 h. CCK-8 assay was used to detect cell viability. C Western blot was used to determine the efficiency of siRNA-mediated knockdown of SLC7A11 in eESCs (50 nM). D–J EESCs underwent indicated treatment: rIL-33(100 ng/ml) and/or transfection with siSLC7A11 (50 nM); GSH (1.5 mM) and/or transfection with siST2 (50 nM). D, E Cell viability was detected by CCK-8 assay. F Western blot was used to detect the protein levels of SLC7A11, and GPX4 in eESCs. G, H Determinations of intracellular MDA (G) and GSH levels (H). I FerroOrange reagent (1 μM) was used to measure intracellular Fe2+concentration. (original magnification ×200). J LiperFluo reagent (5 μM) was used to determine intracellular lipid peroxidation level. (original magnification ×200). Data are presented as the mean ± SD, n = 3 independent experiments. Statistical analysis was performed using Student’s t test (C) or one-way ANOVA (D, E, G, H) or Two-way ANOVA (F). ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05, ns non-significant, siSLC siRNA targeting SLC7A11.