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

Journal: Journal of Radiation Research

Article Title: Increased effectiveness of carbon ions in the production of reactive oxygen species in normal human fibroblasts

doi: 10.1093/jrr/rru083

Figure Lengend Snippet: Distributions of DHE fluorescence intensities (FI; arbitrary units, AU) in AG1522 cells 3 days after X-ray ( A ) or carbon ion ( B ) irradiation with 0.5 Gy. The number of cells as percentage of all cells is shown as a function of the fluorescence intensity.

Article Snippet: For measurements of ROS, cells were trypsinized, washed and resuspended at a concentration of ∼10 5 cells ml –1 in 4 μM Dihydroethidium (DHE; AnaSpec, Fremont, CA, USA) dissolved in PBS.

Techniques: Fluorescence, Irradiation

Journal: Journal of Radiation Research

Article Title: Increased effectiveness of carbon ions in the production of reactive oxygen species in normal human fibroblasts

doi: 10.1093/jrr/rru083

Figure Lengend Snippet: Dose-dependence of ROS levels comparing different fibroblasts (measured by staining with DHE). The fluorescence intensities relative to unirradiated cells (RFI) are shown as a function of the X-ray dose for AG1522 ( n = 6), VH7 and IMR-90 (both n = 1). Plotted are the maximum fluorescence intensities for each dose, typically observed between 3 and 5 days after exposure (see ‘Data analysis’ for details; the corresponding maximum levels can be inferred from Fig. ).

Article Snippet: For measurements of ROS, cells were trypsinized, washed and resuspended at a concentration of ∼10 5 cells ml –1 in 4 μM Dihydroethidium (DHE; AnaSpec, Fremont, CA, USA) dissolved in PBS.

Techniques: Staining, Fluorescence

Journal: Journal of Radiation Research

Article Title: Increased effectiveness of carbon ions in the production of reactive oxygen species in normal human fibroblasts

doi: 10.1093/jrr/rru083

Figure Lengend Snippet: Effect of radiation quality on the dose-dependence of ROS levels in AG1522 cells. RFI (DHE fluorescence intensities relative to unirradiated cells) are shown as a function of dose for carbon ion irradiation ( n = 3) or X-rays ( n = 6, data from Fig. ). The maximum fluorescence intensities for each dose, typically obtained between 3 and 5 days after exposure, were used in the dose–response plot (see ‘Data analysis’ for details). Significant differences between the same physical doses of X-rays and carbon ions; three asterisks indicate P < 0.001.

Article Snippet: For measurements of ROS, cells were trypsinized, washed and resuspended at a concentration of ∼10 5 cells ml –1 in 4 μM Dihydroethidium (DHE; AnaSpec, Fremont, CA, USA) dissolved in PBS.

Techniques: Fluorescence, Irradiation

Journal: Journal of Radiation Research

Article Title: Increased effectiveness of carbon ions in the production of reactive oxygen species in normal human fibroblasts

doi: 10.1093/jrr/rru083

Figure Lengend Snippet: ROS levels and corresponding cell densities of AG1522 cells after X-ray ( A , B ; n = 3–8) or carbon ion irradiation ( C , D ; n = 3) as a function of the time after irradiation. Relative fluorescence intensities (RFIs) of DHE normalized to the mean of unirradiated cells (A, C) are shown. In (B) and (D) the corresponding cell densities are depicted. Significant difference of RFI from 0 Gy is indicated by a single asterisk ( P < 0.05), two asterisks ( P < 0.01) or three asterisks ( P < 0.001). Note that the single experiments are averaged over the timepoints, resulting in error bars and significances that are different from Fig. (see ‘Data analysis’ for details).

Article Snippet: For measurements of ROS, cells were trypsinized, washed and resuspended at a concentration of ∼10 5 cells ml –1 in 4 μM Dihydroethidium (DHE; AnaSpec, Fremont, CA, USA) dissolved in PBS.

Techniques: Irradiation, Fluorescence

Journal: Journal of Radiation Research

Article Title: Increased effectiveness of carbon ions in the production of reactive oxygen species in normal human fibroblasts

doi: 10.1093/jrr/rru083

Figure Lengend Snippet: Representative distributions of DHE ( A ) or p21 ( B ) fluorescence intensities in AG1522 cells 4, 5, 7 and 11 days after irradiation with X-rays (6 Gy, filled histograms; 0 Gy, blank histograms). The number of cells as a percentage of all cells is shown as a function of the fluorescence intensity. ‘X’ indicates an emerging subpopulation that demonstrated reduced p21 or DHE fluorescence intensities.

Article Snippet: For measurements of ROS, cells were trypsinized, washed and resuspended at a concentration of ∼10 5 cells ml –1 in 4 μM Dihydroethidium (DHE; AnaSpec, Fremont, CA, USA) dissolved in PBS.

Techniques: Fluorescence, Irradiation

Journal: Journal of Radiation Research

Article Title: Increased effectiveness of carbon ions in the production of reactive oxygen species in normal human fibroblasts

doi: 10.1093/jrr/rru083

Figure Lengend Snippet: ROS levels in the descendant cells of irradiated AG1522 cells. Shown is the relative fluorescence intensity (RFI) of DHE at an early and a late timepoint after exposure. Filled bars indicate irradiated cells ( n = 7 for 6 Gy X-rays; n = 3 for 2 Gy carbon ions); open bars indicate the descendants ( n = 2 (duplicates), * single measurement) of cells irradiated with 6 Gy X-rays or 2 Gy carbon ions. The dashed horizontal line indicates the level of unirradiated cells at the respective timepoint.

Article Snippet: For measurements of ROS, cells were trypsinized, washed and resuspended at a concentration of ∼10 5 cells ml –1 in 4 μM Dihydroethidium (DHE; AnaSpec, Fremont, CA, USA) dissolved in PBS.

Techniques: Irradiation, Fluorescence

Journal: Theranostics

Article Title: Pseurotin A Inhibits Osteoclastogenesis and Prevents Ovariectomized-Induced Bone Loss by Suppressing Reactive Oxygen Species

doi: 10.7150/thno.30206

Figure Lengend Snippet: Pse treatment reduces osteoclasts and ROS production in OVX mice. (A) Representative images of TRAcP staining of decalcified bone sections. (B-C) Quantitative analyses of N.Oc/BS and Oc.S/B (n=4 per group). (D) Representative images of bone cryosections showing ROS fluorescence in different groups. (E) Quantitative analyses of ROS fluorescence intensity relative to sham group (n=5 per group). All bar graphs are presented as mean ± SD. **P<0.01 relative to the OVX+Vehicle group. BM, bone marrow; DAPI, 4,6-diamidino-2-phenylindole; DHE, dihydroethidium; GP, growth plate; ROS, reactive oxygen species; N.Oc/BS, osteoclast number/bone surface; Oc.S/BS, osteoclast surface/bone surface; TB, trabecular bone; TRAcP, tartrate resistant acid phosphatase

Article Snippet: Dihydroethidium (DHE) was purchased from ApexBio (Boston, MA, USA).

Techniques: Staining, Fluorescence

Journal: Oxidative Medicine and Cellular Longevity

Article Title: In Vitro Evaluation of the Toxicological Profile and Oxidative Stress of Relevant Diet-Related Advanced Glycation End Products and Related 1,2-Dicarbonyls

doi: 10.1155/2021/9912240

Figure Lengend Snippet: Free radical and antioxidant enzymes production upon application of 1,2-dicarbonyls. (a) Production of peroxide and peroxynitrite as measured by DHR fluorescence intensity. (b) Production of superoxide as measured by DHE fluorescence intensity. (c) Production of mitochondrial superoxide as measures by MitoSOX fluorescence intensity. (d) Representative western blot experiment. (e) Production of SOD1 as analyzed by quantification of western blot optical density. (f) Production of SOD2 as analyzed by quantification of western blot optical density. (g) Production of catalase as analyzed by quantification of western blot optical density. (h) Ratio between the production of SOD2 and catalase. Arbitrary 1.0 value was given to the WT group. Here, cell treatments consisted of 750 μ M 3-DGal, 75 μ M 3,4-DGE, 1 mM GO, and 500 μ M MGO. Results are expressed as mean ± SEM ( n = 3). ∗ p < 0.05 vs . CON, ∗∗ p < 0.01 vs . CON, ∗∗∗ p < 0.001 vs . CON.

Article Snippet: To measure O 2 •- production, cells were incubated in 500 μ L 5 μ mol/L dihydroethidium (DHE) (Bioquochem S.L., Llanera, Spain) for 30 minutes at RT.

Techniques: Fluorescence, Western Blot

Journal: Advanced Science

Article Title: Disturbing Cholesterol/Sphingolipid Metabolism by Squalene Epoxidase Arises Crizotinib Hepatotoxicity

doi: 10.1002/advs.202414923

Figure Lengend Snippet: The SQLE enzymatic inhibitor terbinafine interferes with hepatotoxicity of crizotinib. A–F) In vivo model of terbinafine intervention with crizotinib‐induced hepatotoxicity (n = 6 per group). A) The schematic diagram about construction of in vivo model: 1 week period was provided to allow the animals to adapt to the laboratory environment and then C57BL/6J mice were administered 20% cyclodextrin, 100 mg kg −1 crizotinib, 20 mg kg −1 terbinafine or crizotinib plus terbinafine by gavage daily for 6 weeks. Livers and serum were harvested (n = 6 per group). B) Relative expression of c‐PARP was analyzed by western blot with ACTB as a loading control. C) Representative fluorescent image of liver tissues stained with TUNEL and DAPI. White arrows indicated apoptotic cells. The number of TUNEL + cells per field was calculated. D) Serum ALT, AST, LDH, and ALP levels were analyzed. E) Representative images of liver tissues stained with H&E, scale bar = 25 µm. The inflammatory cell infiltration area was analyzed with Fiji software. F) Representative images of liver tissues stained with Oil Red O, scale bar = 50 µm. The Oil Red O‐positive area was analyzed with Fiji software. G) Liver TC and TG levels were analyzed. H) Hepatic NADP + and NADHP levels were measured. I) Representative fluorescent image of liver tissues stained with DHE and DAPI. The relative DHE florescent intensity was analyzed with Fiji software. The results are presented as the mean ± SD. The P value was calculated by one‐way ANOVA (Dunnett's multiple comparisons test). n.s = no significance; * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: For tissue dihydroethidium (DHE) staining, liver tissues were fixed with 4% paraformaldehyde overnight and then dehydrated with 30% sucrose buffer (10 021 418, Sinopharm Chemical Reagent, shanghai, Chian).

Techniques: In Vivo, Expressing, Western Blot, Control, Staining, TUNEL Assay, Software