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2′,7′-dichlorofluorescin diacetate dcfh-da  (Beijing Solarbio Science)
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fluorescent probe 10 μ m 2′,7′-dichlorofluorescein diacetate dcfh-da  (Keygen Biotech)
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Fluorescent Probe 10 μ M 2′,7′ Dichlorofluorescein Diacetate Dcfh Da, supplied by Keygen Biotech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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oxidation-sensitive fluorescent probe dcfh-da kit  (Beyotime)
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peroxide-sensitive fluorescent probe 2′,7′-dichlorofluorescin in diacetate (dcfh-da)  (Genmed Inc)
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dcfh-da reactive oxygen species ros fluorescence probe  (Beijing Solarbio Science)
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Spautin‐1 caused <t>ROS‐mediated</t> DNA damage in melanoma cells. A, Effects of spautin‐1 on ROS generation in A375 and Sk‐Mel‐28 cells. The cells were treated with 10 μmol/L spautin‐1 for 0, 3, 6 or 12 h. Cells were stained with DCF <t>fluorescence</t> probe and the generation of ROS was measured by flow cytometry. The bar graphs showed the relative ROS levels. (Mean values ± SEM, n = 3) * P < .05. B, A375 and Sk‐Mel‐28 were pre‐treated with or without antioxidant NAC (2 mmol/L) for 1 h and then, exposed to spautin‐1 (10 μmol/L) or control medium for another 6 h. Cells were stained with DCF fluorescence probe and the generation of ROS was measured by flow cytometry. The bar graphs showed the relative ROS levels. (Mean values ± SEM, n = 3) * P < .05. C, Cells were treated with spautin‐1 (control, 2.5 or 10 μmol/L) for 48 h. Effects of spautin‐1 on the expression levels of ATM, p‐ATM, ATR, p‐ATR and γ‐H2AX in A375 and Sk‐Mel‐28 cells were examined by Western blot assay. D, A375 and Sk‐Mel‐28 were pre‐treated with or without antioxidant NAC (2 mmol/L) for 1 h and then exposed to spautin‐1 (10 μmol/L) or control medium for another 6 h. Western blot assay was then implied to detect the indicated protein level. E, (Left panel) Representative images of immunofluorescence staining of γ‐H2AX in A375 and Sk‐Mel‐28 cells treated with 10 μmol/L spautin‐1 for 0‐48 h. (Right panel) Quantitative analysis of γ‐H2AX nucleus foci. (Mean values ± SEM, n = 3) Significant differences were evaluated using a one‐way ANOVA. * P < .05 vs control
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dcfh-da fluorescent probe s033s  (Beyotime)
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Spautin‐1 caused <t>ROS‐mediated</t> DNA damage in melanoma cells. A, Effects of spautin‐1 on ROS generation in A375 and Sk‐Mel‐28 cells. The cells were treated with 10 μmol/L spautin‐1 for 0, 3, 6 or 12 h. Cells were stained with DCF <t>fluorescence</t> probe and the generation of ROS was measured by flow cytometry. The bar graphs showed the relative ROS levels. (Mean values ± SEM, n = 3) * P < .05. B, A375 and Sk‐Mel‐28 were pre‐treated with or without antioxidant NAC (2 mmol/L) for 1 h and then, exposed to spautin‐1 (10 μmol/L) or control medium for another 6 h. Cells were stained with DCF fluorescence probe and the generation of ROS was measured by flow cytometry. The bar graphs showed the relative ROS levels. (Mean values ± SEM, n = 3) * P < .05. C, Cells were treated with spautin‐1 (control, 2.5 or 10 μmol/L) for 48 h. Effects of spautin‐1 on the expression levels of ATM, p‐ATM, ATR, p‐ATR and γ‐H2AX in A375 and Sk‐Mel‐28 cells were examined by Western blot assay. D, A375 and Sk‐Mel‐28 were pre‐treated with or without antioxidant NAC (2 mmol/L) for 1 h and then exposed to spautin‐1 (10 μmol/L) or control medium for another 6 h. Western blot assay was then implied to detect the indicated protein level. E, (Left panel) Representative images of immunofluorescence staining of γ‐H2AX in A375 and Sk‐Mel‐28 cells treated with 10 μmol/L spautin‐1 for 0‐48 h. (Right panel) Quantitative analysis of γ‐H2AX nucleus foci. (Mean values ± SEM, n = 3) Significant differences were evaluated using a one‐way ANOVA. * P < .05 vs control
Dcfh Da Fluorescent Probe S033s, supplied by Beyotime, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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dcfh-da fluorescent probe reactive oxygen detection kit  (Biotronik GmbH)
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Biotronik GmbH dcfh-da fluorescent probe reactive oxygen detection kit
Spautin‐1 caused <t>ROS‐mediated</t> DNA damage in melanoma cells. A, Effects of spautin‐1 on ROS generation in A375 and Sk‐Mel‐28 cells. The cells were treated with 10 μmol/L spautin‐1 for 0, 3, 6 or 12 h. Cells were stained with DCF <t>fluorescence</t> probe and the generation of ROS was measured by flow cytometry. The bar graphs showed the relative ROS levels. (Mean values ± SEM, n = 3) * P < .05. B, A375 and Sk‐Mel‐28 were pre‐treated with or without antioxidant NAC (2 mmol/L) for 1 h and then, exposed to spautin‐1 (10 μmol/L) or control medium for another 6 h. Cells were stained with DCF fluorescence probe and the generation of ROS was measured by flow cytometry. The bar graphs showed the relative ROS levels. (Mean values ± SEM, n = 3) * P < .05. C, Cells were treated with spautin‐1 (control, 2.5 or 10 μmol/L) for 48 h. Effects of spautin‐1 on the expression levels of ATM, p‐ATM, ATR, p‐ATR and γ‐H2AX in A375 and Sk‐Mel‐28 cells were examined by Western blot assay. D, A375 and Sk‐Mel‐28 were pre‐treated with or without antioxidant NAC (2 mmol/L) for 1 h and then exposed to spautin‐1 (10 μmol/L) or control medium for another 6 h. Western blot assay was then implied to detect the indicated protein level. E, (Left panel) Representative images of immunofluorescence staining of γ‐H2AX in A375 and Sk‐Mel‐28 cells treated with 10 μmol/L spautin‐1 for 0‐48 h. (Right panel) Quantitative analysis of γ‐H2AX nucleus foci. (Mean values ± SEM, n = 3) Significant differences were evaluated using a one‐way ANOVA. * P < .05 vs control
Dcfh Da Fluorescent Probe Reactive Oxygen Detection Kit, supplied by Biotronik GmbH, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cytoplasmic ros probe fluorescence 2′,7′-dichlorofluorescein diacetate (dcfh-da)  (Beyotime)
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Beyotime cytoplasmic ros probe fluorescence 2′,7′-dichlorofluorescein diacetate (dcfh-da)
Spautin‐1 caused <t>ROS‐mediated</t> DNA damage in melanoma cells. A, Effects of spautin‐1 on ROS generation in A375 and Sk‐Mel‐28 cells. The cells were treated with 10 μmol/L spautin‐1 for 0, 3, 6 or 12 h. Cells were stained with DCF <t>fluorescence</t> probe and the generation of ROS was measured by flow cytometry. The bar graphs showed the relative ROS levels. (Mean values ± SEM, n = 3) * P < .05. B, A375 and Sk‐Mel‐28 were pre‐treated with or without antioxidant NAC (2 mmol/L) for 1 h and then, exposed to spautin‐1 (10 μmol/L) or control medium for another 6 h. Cells were stained with DCF fluorescence probe and the generation of ROS was measured by flow cytometry. The bar graphs showed the relative ROS levels. (Mean values ± SEM, n = 3) * P < .05. C, Cells were treated with spautin‐1 (control, 2.5 or 10 μmol/L) for 48 h. Effects of spautin‐1 on the expression levels of ATM, p‐ATM, ATR, p‐ATR and γ‐H2AX in A375 and Sk‐Mel‐28 cells were examined by Western blot assay. D, A375 and Sk‐Mel‐28 were pre‐treated with or without antioxidant NAC (2 mmol/L) for 1 h and then exposed to spautin‐1 (10 μmol/L) or control medium for another 6 h. Western blot assay was then implied to detect the indicated protein level. E, (Left panel) Representative images of immunofluorescence staining of γ‐H2AX in A375 and Sk‐Mel‐28 cells treated with 10 μmol/L spautin‐1 for 0‐48 h. (Right panel) Quantitative analysis of γ‐H2AX nucleus foci. (Mean values ± SEM, n = 3) Significant differences were evaluated using a one‐way ANOVA. * P < .05 vs control
Cytoplasmic Ros Probe Fluorescence 2′,7′ Dichlorofluorescein Diacetate (Dcfh Da), supplied by Beyotime, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Spautin‐1 caused ROS‐mediated DNA damage in melanoma cells. A, Effects of spautin‐1 on ROS generation in A375 and Sk‐Mel‐28 cells. The cells were treated with 10 μmol/L spautin‐1 for 0, 3, 6 or 12 h. Cells were stained with DCF fluorescence probe and the generation of ROS was measured by flow cytometry. The bar graphs showed the relative ROS levels. (Mean values ± SEM, n = 3) * P < .05. B, A375 and Sk‐Mel‐28 were pre‐treated with or without antioxidant NAC (2 mmol/L) for 1 h and then, exposed to spautin‐1 (10 μmol/L) or control medium for another 6 h. Cells were stained with DCF fluorescence probe and the generation of ROS was measured by flow cytometry. The bar graphs showed the relative ROS levels. (Mean values ± SEM, n = 3) * P < .05. C, Cells were treated with spautin‐1 (control, 2.5 or 10 μmol/L) for 48 h. Effects of spautin‐1 on the expression levels of ATM, p‐ATM, ATR, p‐ATR and γ‐H2AX in A375 and Sk‐Mel‐28 cells were examined by Western blot assay. D, A375 and Sk‐Mel‐28 were pre‐treated with or without antioxidant NAC (2 mmol/L) for 1 h and then exposed to spautin‐1 (10 μmol/L) or control medium for another 6 h. Western blot assay was then implied to detect the indicated protein level. E, (Left panel) Representative images of immunofluorescence staining of γ‐H2AX in A375 and Sk‐Mel‐28 cells treated with 10 μmol/L spautin‐1 for 0‐48 h. (Right panel) Quantitative analysis of γ‐H2AX nucleus foci. (Mean values ± SEM, n = 3) Significant differences were evaluated using a one‐way ANOVA. * P < .05 vs control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Potent USP10/13 antagonist spautin‐1 suppresses melanoma growth via ROS‐mediated DNA damage and exhibits synergy with cisplatin

doi: 10.1111/jcmm.15093

Figure Lengend Snippet: Spautin‐1 caused ROS‐mediated DNA damage in melanoma cells. A, Effects of spautin‐1 on ROS generation in A375 and Sk‐Mel‐28 cells. The cells were treated with 10 μmol/L spautin‐1 for 0, 3, 6 or 12 h. Cells were stained with DCF fluorescence probe and the generation of ROS was measured by flow cytometry. The bar graphs showed the relative ROS levels. (Mean values ± SEM, n = 3) * P < .05. B, A375 and Sk‐Mel‐28 were pre‐treated with or without antioxidant NAC (2 mmol/L) for 1 h and then, exposed to spautin‐1 (10 μmol/L) or control medium for another 6 h. Cells were stained with DCF fluorescence probe and the generation of ROS was measured by flow cytometry. The bar graphs showed the relative ROS levels. (Mean values ± SEM, n = 3) * P < .05. C, Cells were treated with spautin‐1 (control, 2.5 or 10 μmol/L) for 48 h. Effects of spautin‐1 on the expression levels of ATM, p‐ATM, ATR, p‐ATR and γ‐H2AX in A375 and Sk‐Mel‐28 cells were examined by Western blot assay. D, A375 and Sk‐Mel‐28 were pre‐treated with or without antioxidant NAC (2 mmol/L) for 1 h and then exposed to spautin‐1 (10 μmol/L) or control medium for another 6 h. Western blot assay was then implied to detect the indicated protein level. E, (Left panel) Representative images of immunofluorescence staining of γ‐H2AX in A375 and Sk‐Mel‐28 cells treated with 10 μmol/L spautin‐1 for 0‐48 h. (Right panel) Quantitative analysis of γ‐H2AX nucleus foci. (Mean values ± SEM, n = 3) Significant differences were evaluated using a one‐way ANOVA. * P < .05 vs control

Article Snippet: The effect of spautin‐1 on intracellular ROS generation was measured using a DCFH‐DA reactive oxygen species ROS fluorescence probe (Solarbio).

Techniques: Staining, Fluorescence, Flow Cytometry, Control, Expressing, Western Blot, Immunofluorescence