Journal: Redox Biology
Article Title: USP20 governs tyrosine kinase inhibitors resistance through ferroptosis evasion by targeting GPX4 in cancers
doi: 10.1016/j.redox.2026.104086
Figure Lengend Snippet: USP20 elevates GPX4 protein abundance through post-translational regulation. ( A ) Western blot analysis of GPX4 and USP20 proteins in 769-P and A549 cells transfected with lentivirus carrying shUSP20 or negative control. ( B ) Western blot analysis of GPX4 and USP20 proteins in 786-O and HEK 293T cells transfected with GST-USP20 plasmids or vector. ( C-D ) Analysis of GPX4 and USP20 mRNA expression in shScr and shUSP20 769-P and A549 cells. ( E-H ) shScr and shUSP20 769-P (E-F) and A549 (G-H) cells were treated with 100 μg/mL CHX at indicated time points. The GPX4 protein abundance was quantified by the Image J and statistical charts are produced by GraphPad. ( I-J ) shScr and shUSP20 769-P and A549 cells were treated with 20 μM MG132 for 6 h or 20 μM CQ for 12 h, and then assessed GPX4 expression. ( K ) Western blot analysis of GPX4 proteins in HEK293 cells transfected with GST-USP20 WT or GST-USP20 C154S plasmids. ( L-M ) Western blot analysis of GPX4 proteins in USP20 knockdown HEK293 cells transfected with GST-Vector, GST-USP20 WT or GST-USP20 C154S plasmids. The GPX4 protein abundance was quantified by the Image J software and statistical charts are produced by GraphPad. ( N ) Western blot analysis of GPX4 proteins in 769-P, SW839, A549, H1299 cells treated with indicated concentration GSK2643943A for 24 h. ( O–R ) The 769-P and A549 cells were treated with 10 μM GSK2643943A for 24 h, then 100 μg/mL CHX at indicated time points. The GPX4 protein abundance was quantified by the Image J software and statistical charts are produced by GraphPad. Data in C and D are presented as mean ± s.d. of n = 3 biological replicates.
Article Snippet: Human RCC cell lines 769-P, 786-O and SW839, human NSCLC cell lines A549, H1299 and the HEK-293T cell line were obtained from the American Type Culture Collection and cultured in RPMI-1640 or DMEM medium (Thermo Fisher Scientific, Inc.) added with 10% fetal bovine serum (FBS) (Thermo Fisher Scientific, Inc.),100 U/ml penicillin and 0.1 mg/ml streptomycin (Thermo Fisher Scientific, Inc.) at a temperature of 37 °C.
Techniques: Quantitative Proteomics, Western Blot, Transfection, Negative Control, Plasmid Preparation, Expressing, Produced, Knockdown, Software, Concentration Assay
