dq bsa  (Tocris)

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 rescues lysosomal dysfunction and promotes autophagosome–lysosome fusion under DOX-induced stress. Cells were transduced with Ad-PrxⅢ or Ad-Stuffer for 24 h and then exposed to 1 μM DOX for 6 h. (A) Lysosomal protease activity was analyzed using Magic Red (red), and nuclei were counterstained with Hoechst (blue). Scale bar, 25 μm. (B, C) Lysosomal acidity was analyzed by (B) DQ-BSA Red (red; scale bar, 10 μm) and (C) LysoSensor Green DND-189 (green; scale bar, 25 μm). Nuclei were stained with DAPI (blue). (D) Cells were transduced with Ad-PrxⅢ or Ad-Stuffer in the presence of GFP-LC3 (green) adenovirus for 24 h and treated as above. Fixed cells were immunostained with anti-LAMP1 antibody (red), and nuclei were stained with DAPI (blue). Colocalization of GFP-LC3 and LAMP1 was plotted as yellow dots in graph. Scale bar, 15 μm. Five randomly selected microscopic fields per sample were analyzed and combined to represent one biological replicate. All data are presented as mean ± S.D. Statistical significance was determined using two-way ANOVA followed by Bonferroni's post hoc test. ∗∗p < 0.01 and ∗∗∗p < 0.001.

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, Activity Assay, Staining