Fig. 5
Catalytically inactive PRDX1 variants, in contrast to C83A mutant and wtPRDX1 variant, do not rescue survival of PRDX1-knockout MCF-7 cells. a anti-PRDX1 and anti-V5-tag western blotting analysis shows overexpression of PRDX1 mutated protein variants with the following mutations: C83A, C173A, and C52/C173A (lanes 3–5, respectively) or overexpressing wtPRDX1 protein (lane 6) in MCF-7 sgPRDX1-A cells compared to parental (lane 1) and knockout sgPRDX1-A (lane 2) cells. β-actin was used as a loading control. b, c Cells were treated with increasing concentrations of glucose oxidase (0.125–2 mU) (b) or sodium L-ascorbate (0.1–1.6 mM) (c) for 24 h. Catalytically inactive PRDX1 variants, in contrast to C83A mutant and wtPRDX1 variant, do not rescue survival of PRDX1-knockout MCF-7 cells. For all cytotoxicity assays, control cells were cultured without any reagent. At the end of treatment, the crystal violet staining was performed and reported as percent growth relative to control. Experiments were performed in triplicates and repeated at least twice. Statistical analysis was performed with one-way ANOVA followed by Tukey’s honestly significant difference (HSD) post hoc test when significance was detected (*p < 0.05, **p < 0.01, ***p < 0.001)
