Fig. 2

Juglone induces cell death phenotypic changes in GBM cells. A LN229 and T98G cells were pretreated with inhibitors such as Fer-1 (4 μM), Lip-1 (0.1 μM), Nec-1 (10 μM), Z-VAD (25 μM), CQ-1(10 μM) or Mdivi(20 μM) for 1 h and then co-treated with juglone for 24 h. The cell viability was determined through a CCK-8 assay. B, C LN229 and T98G cells were treated with different concentrations of juglone for 24 h. The ROS levels of each group of cells were detected and quantitatively analyzed through flow cytometry. D, E The levels of GSH and MDA in LN229 and T98G cells were measured under the effect of different concentrations of juglone. F, G After 24 h of treatment with different concentrations of juglone, the 4HNE level in LN229 and T98G cells was detected and quantitatively analyzed through immunofluorescence. Fer-1 ferrostatin-1, Lip-1 liproxstatin-1, Nec necrostatin-1, Z-VAD Z-VAD-FMK, CQ-1 chloroquine, Mdivi Mdivi-1, GSH glutathione, MDA malonaldehyde; *p < 0.05; **p < 0.01; ***p < 0.001; ns, no statistical significance