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Fig. 5 | Journal of Experimental & Clinical Cancer Research

Fig. 5

From: PRX1 knockdown potentiates vitamin K3 toxicity in cancer cells: a potential new therapeutic perspective for an old drug

Fig. 5

Oxidation of HyPer, endogenous TRX1, TRX2 and PRX3 monitored by redox western blot. a Oxidation of HyPer in response to vitK3 treatment. Cells expressing respectively cyto-, nuc- and mito-HyPer were treated with 15 μM vitK3 for indicated times and then subjected to redox western blots. Quantifications are mean ± SD of at least three independent experiments. b Redox western blot for TRX1 and TRX2. Cells treated with 15 μM vitK3 for indicated times were subjected to redox protein extraction and AMS treatment. Lane PC presents TRX1 oxidation in H2O2-treated Prx1+ cells as positive control. Quantifications are mean ± SD of at least three independent experiments. c Redox western blot for PRX3. Redox protein extraction was treated with NEM. Monomer and dimer forms are indicated. d NAC suppresses vitK3 toxicity. Cells were pretreated with 10 mM NAC for 2 h prior to incubation with freshly prepared medium containing both 10 mM NAC and 15 μM vitK3 for additional 4 h. Cell viability was determined by MTT assay (upper panel). e NAC suppresses vitK3-induced ROS accumulation. Global ROS levels were monitored by carboxy-H2DCFDA (left panel). Gray shade, black solid line, and black dotted line indicate the fluorescence intensity of the control cells, cells treated with 15 μM vitK3 alone for 4 h, and cells treated with 10 mM NAC and 15 μM vitK3 for 4 h, respectively. Compartment-specific H2O2 levels in Prx1– cells were monitored by HyPer probes (right panel). Redox state of HyPer in Prx1– cells treated with 15 μM vitK3 for 60 min was used as positive control. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001

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