Journal: Circulation Research

Article Title: Selective Activation of Lectin-Like Oxidized Low-Density Lipoprotein Receptor-1 Mediates C-Reactive Protein–Evoked Endothelial Vasodilator Dysfunction in Coronary Arterioles

doi: 10.1161/circresaha.114.301763

Figure Lengend Snippet: Figure 4. Dihydroethidium (DHE) imaging of superoxide in pressurized coronary arterioles. A, Representative fluorescence images of isolated coronary arterioles with basal tone incubated for 60 minutes in the presence of endothelium with vehicle (control), 7 μg/mL C-reactive protein (CRP), CRP+4-hydroxy- 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPOL) (1 mmol/L), CRP+anti–lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) antibody (5 μg/mL), or CRP+anti-CD32 antibody (5 μg/mL) or in the absence of endothelium (denudation) with CRP, followed by addition of oxidative fluorescent dye DHE for 5 minutes. Scale bar, 50 μm. B, Quantitative analysis of DHE fluorescence signals for control and CRP-treated vessel groups shown in A. Data represent 4 to 5 independent experiments. *P<0.05 vs control. Ab indicates antibody.

Article Snippet: Real-time detection of superoxide production from isolated porcine coronary arterioles in the presence or absence of endothelium was conducted using dihydroethidium (DHE) with slight modification from a previous study.16 Isolated and pressurized coronary arterioles were incubated intraluminally with PSS containing vehicle, CRP (7 μg/mL), CRP plus TEMPOL (1 mmol/L), CRP plus anti-LOX-1 antibody (5 μg/mL, Abcam), or CRP plus anti-CD32 antibody (5 μg/mL, Santa Cruz Biotechnology) at 37°C for 60 minutes.

Techniques: Imaging, Fluorescence, Isolation, Incubation, Control

Journal: Frontiers in Pharmacology

Article Title: Natural compound Alternol exerts a broad anti-cancer spectrum and a superior therapeutic safety index in vivo

doi: 10.3389/fphar.2024.1409506

Figure Lengend Snippet: Detection of ROS probe distribution after Alternol treatment. RMG-I and OVSAHO cells were seeded in 6-well plates and treated with the solvent or Alternol (5 μM) for 4 h. After washing in PBS, cells were incubated with the fluorescent probes (5 μM), MitoSOX panel (A) , DHE panel (B) , or PFBS panel (C) , for 30 min followed by flow cytometer analysis of probe distribution.

Article Snippet: The dihydroethidium (DHE)-based ROS detection assay kit (catalog #601290), propidium iodide (Catalog #14289), and penta-fluoro-benzene-sulfonyl fluorescein (PFBS, Catalog #10005983) were purchased from Cayman Chemicals.

Techniques: Solvent, Incubation, Flow Cytometry

Journal: International Journal of Nanomedicine

Article Title: Fe-HCOF-PEG 2000 as a Hypoxia-Tolerant Photosensitizer to Trigger Ferroptosis and Enhance ROS-Based Cancer Therapy

doi: 10.2147/IJN.S479848

Figure Lengend Snippet: ROS generation of Fe-HCOF-PEG 2000 . The EPR spectrogram of ( a ) O 2 •− , ( b ) OH• and ( c ) 1 O 2 captured by DMPO or TEMPO when they born by Fe-HCOFs under NIR treatment at 10min or 0min time point. ( d ) Schematic diagram of type I PDT. Fluorescence spectrum of ( e ) DHE and ( f ) DCFH-DA motivated by Fe-HCOFs under different duration of 808nm NIR irradiation.

Article Snippet: Dihydroethidium (Hydroethidine) and DCFH-DA (H2DCFDA) were purchased from Glpbio.

Techniques: Fluorescence, Irradiation

Journal: International Journal of Nanomedicine

Article Title: Fe-HCOF-PEG 2000 as a Hypoxia-Tolerant Photosensitizer to Trigger Ferroptosis and Enhance ROS-Based Cancer Therapy

doi: 10.2147/IJN.S479848

Figure Lengend Snippet: ROS production of Fe-HCOF-PEG 2000 in cancer cells. ( a ) Fluorescence images of DCFH-DA probe bonded 4T1 cells to detect cellular ROS generation. Scale bar: 50 μm. ( b ) Flow cytometry of DCFH-DA probe bonded 4T1 cells to detect cellular ROS generation. ( c and d ) Quantitative intensity of DCFH-DA in Figure a and DHE in Figure e, respectively. n=3. Data are shown as mean ± SD. Student’s t‐test, * p < 0.05, ** p < 0.01, *** p < 0.001. ( e ) DHE staining images of 4T1 cells treated with 5 definite methods to indicate O 2 •− positive cells. Scale bar: 20 μm.

Article Snippet: Dihydroethidium (Hydroethidine) and DCFH-DA (H2DCFDA) were purchased from Glpbio.

Techniques: Fluorescence, Flow Cytometry, Staining