Journal: JCI Insight
Article Title: 4EBP1-mediated SLC7A11 protein synthesis restrains ferroptosis triggered by MEK inhibitors in advanced ovarian cancer
doi: 10.1172/jci.insight.177857
Figure Lengend Snippet: ( A ) Cell viability of SKOV3 treated with 500 nM trametinib with or without PI3K/AKT/mTOR inhibitors for 96 hours. BY719 (1 μM), PI103 (1 μM), MK2206 (MK) (5 μM), GSK690693 (GSK) (5 μM), rapamycin (Rapa) (1 μM), everolimus (Evero) (0.5 μM). ( B ) Colony formation of SKOV3 treated with 500 nM trametinib with or without PI3K/AKT/mTOR inhibitors. BY719 (1 μM), PI103 (1 μM), MK2206 (MK) (5 μM), GSK690693 (GSK) (5 μM), rapamycin (Rapa) (1 μM), everolimus (Evero) (0.5 μM). ( C ) Growth curves and ( D ) sub-G 1 population analysis in SKOV3 treated with vehicle, trametinib, AKT inhibitors (GSK690693 or MK2206), or their combination. ( E ) Colony formation assay of PDC-POVC15 treated with trametinib (100 nM) with or without AKT inhibitors (MK2206, 5 μM and GSK690693, 5 μM). ( F ) Cell viability of SKOV3 and A2780R cells following trametinib (500 nM) or MK2206 treatment (5 μM) in the presence or absence of Fer-1 (2 μM), Lipro-1 (100 nM), DFO (300 nM), Z-VAD (5 μM), and Necro-1 (5 μM) for 48 hours. ( G ) Detection of lipid peroxidation level with BODIPY 581/591 C11 probe determined by the flow cytometer in SKOV3 and A2780R treated with trametinib (500 nM) or MK2206 (5 μM) treatment in the presence or absence of Fer-1, Lipro-1, and DFO for 48 hours. ( H ) Detection of GSH level in SKOV3 and A2780R followed by trametinib (500 nM) or MK2206 treatment (5 μM) in the presence or absence of Fer-1 or Lipro-1 for 48 hours. The data are presented as the mean ± SD of 3 independent experiments. ( A and F – H ) P values were determined by 1-way ANOVA with Bonferroni’s post hoc test. ( C ) P values were determined by 2-way ANOVA with Tukey’s post hoc test. ( D ) P values were determined by unpaired Student’s t test. * P < 0.05, ** P < 0.01, *** P < 0.001.
Article Snippet: Ferrostatin-1 (S7243), Liproxstatin-1 (S7699), Z-VAD-FMK (S7023), Necrostatin-1 (S8037), MG132 (S2619), PI103 (S1038), BY719 (S2814), rapamycin (S1039), everolimus (S1120), erastin (S7242), and kinase inhibitor drug library were purchased from Selleckchem.com.
Techniques: Colony Assay, Flow Cytometry
