Figure 3. Inhibition of GSH synthesis induces ferroptosis in ccRCC cells.
A) Diagram illustrating the mechanisms of GSH synthesis and induction of ferroptosis.
B) RCC4 and 786-O cells were treated with the indicated concentrations of Erastin or solvent (DMSO) either in the presence or absence of ferrostatin (fer-1) for 72h. Cell viability was determined by crystal violet staining. Values represent mean ±SEM relative to solvent-treated controls (n=3).
C) RCC4 and 786-O cells were treated with the indicated concentrations of BSO or solvent (DMSO) either in the presence or absence of ferrostatin (fer-1) for 72h. Cell viability was determined by crystal violet staining. Values represent mean ±SEM relative to solvent-treated controls (n=3).
D) RCC4 cells were transfected with siRNA oligonucleotides targeting SLC7A11, UBB or non-targeting controls (CTRL) and treated with 4 μM ferrostatin (fer-1) or solvent for 96 h. Cell viability was determined by crystal violet staining. Values represent mean ±SEM relative to solvent-treated controls (n=3).
E) RCC4 cells were treated with the indicated concentrations of BSO or solvent (DMSO) for 24h. GSH levels were determined by mass spectrometry. Values represent mean ±SEM (n=2).
F) RCC4 and 786-O cells were treated with the indicated concentrations of BSO or solvent (DMSO) with or without different concentrations of hydrogen peroxide (H2O2) for 72 hours. Cell viability was determined by crystal violet staining. Values represent mean ±SEM relative to solvent-treated controls (n=3).
*p≤0.05; **p≤0.01; ***p≤0.005; ****p≤0.001 unpaired two-tailed Student’s t-test.