Journal: Redox Biology

Article Title: Peroxiredoxin Ⅲ safeguards cardiac function against doxorubicin by regulating mitochondrial quality control via H 2 O 2 detoxification

doi: 10.1016/j.redox.2026.104176

Figure Lengend Snippet: PrxⅢ regulates oxidative damage and mitochondrial bioenergetics in DOX-treated cardiomyocytes. (A) Cells were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 12 h. Cells were labeled with 5 μM 10-NAO and relative fluorescence intensity (RFI) was analyzed by flow cytometer. Representative histograms and quantification are shown. (B) Cells were treated with 1 μM DOX for 12 h and labeled with 10 μM Rho-123 to assess mitochondrial membrane potential ( ΔΨ m ). The percentage of cells with low ΔΨ m was quantified by flow cytometry. Representative histograms and quantification are shown. (C) Cells were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 12 h. Representative bar graph showing relative ATP concentration normalized to protein contents. (D-H) Cells were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 12 h. (D) Representative tracing of the oxygen consumption rate (OCR). Arrows indicate time points when cells were treated with oligomycin, FCCP, and rotenone plus antimycin A (Rot/AA), respectively. The OCR is calculated as (E) basal respiration, (F) maximal respiration, (G) spare respiration, and (H) proton-leak. All data are expressed as mean ± S.D. (n = 3-5). Statistical significance was determined using two-way ANOVA followed by Bonferroni's post hoc test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001.

Article Snippet: DOX was kindly provided from Donga-ST Co., Ltd. (Seoul, Republic of Korea); Fetal Bovine Serum (FBS) and antibiotic-antimycotic (A/A) were from Gibco (NY, USA); Dimethyl sulfoxide (DMSO), poly- l -lysine, puromycin, and bafilomycin A1 (BafA1) were from Sigma-Aldrich (St. Louis, MO, USA); 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium monosodium salt (WST-1) reagent was purchased from Roche Applied Science (Basel, Switzerland); Lipofectamine 2000, 5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate (CM-H2DCFDA), Rhodamine-123 (Rho-123), Mitotracker Red CMX, Lysosensor Green DND-189, DQ-BSA and 7-Aminoactinomycin D (7-AAD) were purchased from Invitrogen (CA, USA); 10-N-nonylacridine orange (10-NAO) was purchased from Molecular Probes (Eugene, OR, USA); Mitochondria peroxy yellow-1 (MitoPY-1) was from Tocris Biosciences (Bristol, UK); Magic Red kit was from ImmunoChemistry Technologies (Davis, CA, USA); paraformaldehyde (PFA), 4′,6-diamidno-2-phenylindole (DAPI) were from Thermo Fisher Scientific (Waltham, MA, USA); Adenosine triphosphate (ATP) and cell viability ATP assay kit was from Promega (Madison, WI, USA); Seahorse XF cell Mitostress test kit, Seahorse XF DMEM Medium, and Seahorse XF Calibrant Solution was from Agilent (Santa Clara, CA, USA); Elamipretide (SS-31) was from Biorbyt Ltd. (Cambridge, UK); Mitoquinone (MitoQ) and Visomitin (SKQ1) were from MedChemExpress (Monmouth junction, NJ, USA); Mouse Troponin I ELISA kit was from Abcam (Cambridge, UK).

Techniques: Transduction, Labeling, Fluorescence, Flow Cytometry, Membrane, Concentration Assay