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Cell Signaling Technology Inc prolong gold antifade reagent with dapi
Fig. 3 CX-5461 causes DNA damage and cell death by apoptosis in neuroblastoma cells at sub-micromolar concentrations. a Apoptosis in CHP-134 cells. The cells were treated for 48 h. b Quantification of data in (a). Data represent the mean ± SD of three independent experiments. Significant P-values (left to right): 3.9 × 10−6, 7.2 × 10−7, and 4.8 × 10−7. c Quantification of flow cytometry data showing apoptosis up to 48 h in CHP-134 and IMR-5 (0.2 µM and 0.1 µM of CX-5461 respectively). CHP-134 cells were treated with 0.2 µM and IMR-5 cells with 0.1 µM of CX-5461. Data represent the mean ± SD of three independent experiments. Significant P-values (left to right): CHP-134, 2.2 × 10−4, 3.2 × 10−5, and 1.5 × 10−5; IMR-5, 1.8 × 10−4, 2.3 × 10−5, and 2.5 × 10−6. d Western blots for cleaved PARP (c-PARP). β-Actin was used as a loading control. Representative results of three biological replicates are shown. e Co-localization of 53BP1 and γ-H2AX. Cells were treated for 24 h. Representative results of three biological replicates are shown. Scale bar = 10 μm. See Fig. S3d for more time points. f Comet assay in CHP-134 cells. Tail length (left) and representative images (right) are shown. The red line indicates the mean value of tail length in each group. Scale bar = 5 µm. g Cell cycle analysis forCHP-134 (0.2 µM CX-5461) and IMR-5 (0.05 µM CX-5461) cells. Data represent the mean ± SD of three independent experiments. P-values vs 0 h: P1 = 4.2 × 10−5, P2 = 7.1 × 10−7, P3 = 6.3 × 10−5, P4 = 2.7 × 10−4, P5 = 6.3 × 10−3, P6 = 2.3 × 10−5, P7 = 1.8 × 10−4, and P8 = 0.044. h EdU incorporation assay to assess global nascent DNA transcription. Representative results of three biological replicates are shown. Blue, <t>DAPI;</t> green, EdU. Scale bar = 20 µm. i CX-5461 induced DNA damage (γ-H2AX, red) was enriched in cells with active DNA synthesis (positive for EdU (green) labeling). Representative results of three biological replicates are shown. Scale bar = 10 μm. Two-tailed t- test was used for (b, c, g). Source data for these panels are included in Source Data file.
Prolong Gold Antifade Reagent With Dapi, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/prolong gold antifade reagent with dapi/product/Cell Signaling Technology Inc
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Servicebio Inc prolong gold antifade containing dapi gb012
Fig. 3 CX-5461 causes DNA damage and cell death by apoptosis in neuroblastoma cells at sub-micromolar concentrations. a Apoptosis in CHP-134 cells. The cells were treated for 48 h. b Quantification of data in (a). Data represent the mean ± SD of three independent experiments. Significant P-values (left to right): 3.9 × 10−6, 7.2 × 10−7, and 4.8 × 10−7. c Quantification of flow cytometry data showing apoptosis up to 48 h in CHP-134 and IMR-5 (0.2 µM and 0.1 µM of CX-5461 respectively). CHP-134 cells were treated with 0.2 µM and IMR-5 cells with 0.1 µM of CX-5461. Data represent the mean ± SD of three independent experiments. Significant P-values (left to right): CHP-134, 2.2 × 10−4, 3.2 × 10−5, and 1.5 × 10−5; IMR-5, 1.8 × 10−4, 2.3 × 10−5, and 2.5 × 10−6. d Western blots for cleaved PARP (c-PARP). β-Actin was used as a loading control. Representative results of three biological replicates are shown. e Co-localization of 53BP1 and γ-H2AX. Cells were treated for 24 h. Representative results of three biological replicates are shown. Scale bar = 10 μm. See Fig. S3d for more time points. f Comet assay in CHP-134 cells. Tail length (left) and representative images (right) are shown. The red line indicates the mean value of tail length in each group. Scale bar = 5 µm. g Cell cycle analysis forCHP-134 (0.2 µM CX-5461) and IMR-5 (0.05 µM CX-5461) cells. Data represent the mean ± SD of three independent experiments. P-values vs 0 h: P1 = 4.2 × 10−5, P2 = 7.1 × 10−7, P3 = 6.3 × 10−5, P4 = 2.7 × 10−4, P5 = 6.3 × 10−3, P6 = 2.3 × 10−5, P7 = 1.8 × 10−4, and P8 = 0.044. h EdU incorporation assay to assess global nascent DNA transcription. Representative results of three biological replicates are shown. Blue, <t>DAPI;</t> green, EdU. Scale bar = 20 µm. i CX-5461 induced DNA damage (γ-H2AX, red) was enriched in cells with active DNA synthesis (positive for EdU (green) labeling). Representative results of three biological replicates are shown. Scale bar = 10 μm. Two-tailed t- test was used for (b, c, g). Source data for these panels are included in Source Data file.
Prolong Gold Antifade Containing Dapi Gb012, supplied by Servicebio Inc, 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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Promega prolong anti-fade
Fig. 3 CX-5461 causes DNA damage and cell death by apoptosis in neuroblastoma cells at sub-micromolar concentrations. a Apoptosis in CHP-134 cells. The cells were treated for 48 h. b Quantification of data in (a). Data represent the mean ± SD of three independent experiments. Significant P-values (left to right): 3.9 × 10−6, 7.2 × 10−7, and 4.8 × 10−7. c Quantification of flow cytometry data showing apoptosis up to 48 h in CHP-134 and IMR-5 (0.2 µM and 0.1 µM of CX-5461 respectively). CHP-134 cells were treated with 0.2 µM and IMR-5 cells with 0.1 µM of CX-5461. Data represent the mean ± SD of three independent experiments. Significant P-values (left to right): CHP-134, 2.2 × 10−4, 3.2 × 10−5, and 1.5 × 10−5; IMR-5, 1.8 × 10−4, 2.3 × 10−5, and 2.5 × 10−6. d Western blots for cleaved PARP (c-PARP). β-Actin was used as a loading control. Representative results of three biological replicates are shown. e Co-localization of 53BP1 and γ-H2AX. Cells were treated for 24 h. Representative results of three biological replicates are shown. Scale bar = 10 μm. See Fig. S3d for more time points. f Comet assay in CHP-134 cells. Tail length (left) and representative images (right) are shown. The red line indicates the mean value of tail length in each group. Scale bar = 5 µm. g Cell cycle analysis forCHP-134 (0.2 µM CX-5461) and IMR-5 (0.05 µM CX-5461) cells. Data represent the mean ± SD of three independent experiments. P-values vs 0 h: P1 = 4.2 × 10−5, P2 = 7.1 × 10−7, P3 = 6.3 × 10−5, P4 = 2.7 × 10−4, P5 = 6.3 × 10−3, P6 = 2.3 × 10−5, P7 = 1.8 × 10−4, and P8 = 0.044. h EdU incorporation assay to assess global nascent DNA transcription. Representative results of three biological replicates are shown. Blue, <t>DAPI;</t> green, EdU. Scale bar = 20 µm. i CX-5461 induced DNA damage (γ-H2AX, red) was enriched in cells with active DNA synthesis (positive for EdU (green) labeling). Representative results of three biological replicates are shown. Scale bar = 10 μm. Two-tailed t- test was used for (b, c, g). Source data for these panels are included in Source Data file.
Prolong Anti Fade, supplied by Promega, 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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Beijing Solarbio Science prolonged gold anti-fade with dapi
Fig. 3 CX-5461 causes DNA damage and cell death by apoptosis in neuroblastoma cells at sub-micromolar concentrations. a Apoptosis in CHP-134 cells. The cells were treated for 48 h. b Quantification of data in (a). Data represent the mean ± SD of three independent experiments. Significant P-values (left to right): 3.9 × 10−6, 7.2 × 10−7, and 4.8 × 10−7. c Quantification of flow cytometry data showing apoptosis up to 48 h in CHP-134 and IMR-5 (0.2 µM and 0.1 µM of CX-5461 respectively). CHP-134 cells were treated with 0.2 µM and IMR-5 cells with 0.1 µM of CX-5461. Data represent the mean ± SD of three independent experiments. Significant P-values (left to right): CHP-134, 2.2 × 10−4, 3.2 × 10−5, and 1.5 × 10−5; IMR-5, 1.8 × 10−4, 2.3 × 10−5, and 2.5 × 10−6. d Western blots for cleaved PARP (c-PARP). β-Actin was used as a loading control. Representative results of three biological replicates are shown. e Co-localization of 53BP1 and γ-H2AX. Cells were treated for 24 h. Representative results of three biological replicates are shown. Scale bar = 10 μm. See Fig. S3d for more time points. f Comet assay in CHP-134 cells. Tail length (left) and representative images (right) are shown. The red line indicates the mean value of tail length in each group. Scale bar = 5 µm. g Cell cycle analysis forCHP-134 (0.2 µM CX-5461) and IMR-5 (0.05 µM CX-5461) cells. Data represent the mean ± SD of three independent experiments. P-values vs 0 h: P1 = 4.2 × 10−5, P2 = 7.1 × 10−7, P3 = 6.3 × 10−5, P4 = 2.7 × 10−4, P5 = 6.3 × 10−3, P6 = 2.3 × 10−5, P7 = 1.8 × 10−4, and P8 = 0.044. h EdU incorporation assay to assess global nascent DNA transcription. Representative results of three biological replicates are shown. Blue, <t>DAPI;</t> green, EdU. Scale bar = 20 µm. i CX-5461 induced DNA damage (γ-H2AX, red) was enriched in cells with active DNA synthesis (positive for EdU (green) labeling). Representative results of three biological replicates are shown. Scale bar = 10 μm. Two-tailed t- test was used for (b, c, g). Source data for these panels are included in Source Data file.
Prolonged Gold Anti Fade With Dapi, supplied by Beijing Solarbio Science, 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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Diamon Gmbh prolong diamon antifade mountant with dapi reagent
Fig. 3 CX-5461 causes DNA damage and cell death by apoptosis in neuroblastoma cells at sub-micromolar concentrations. a Apoptosis in CHP-134 cells. The cells were treated for 48 h. b Quantification of data in (a). Data represent the mean ± SD of three independent experiments. Significant P-values (left to right): 3.9 × 10−6, 7.2 × 10−7, and 4.8 × 10−7. c Quantification of flow cytometry data showing apoptosis up to 48 h in CHP-134 and IMR-5 (0.2 µM and 0.1 µM of CX-5461 respectively). CHP-134 cells were treated with 0.2 µM and IMR-5 cells with 0.1 µM of CX-5461. Data represent the mean ± SD of three independent experiments. Significant P-values (left to right): CHP-134, 2.2 × 10−4, 3.2 × 10−5, and 1.5 × 10−5; IMR-5, 1.8 × 10−4, 2.3 × 10−5, and 2.5 × 10−6. d Western blots for cleaved PARP (c-PARP). β-Actin was used as a loading control. Representative results of three biological replicates are shown. e Co-localization of 53BP1 and γ-H2AX. Cells were treated for 24 h. Representative results of three biological replicates are shown. Scale bar = 10 μm. See Fig. S3d for more time points. f Comet assay in CHP-134 cells. Tail length (left) and representative images (right) are shown. The red line indicates the mean value of tail length in each group. Scale bar = 5 µm. g Cell cycle analysis forCHP-134 (0.2 µM CX-5461) and IMR-5 (0.05 µM CX-5461) cells. Data represent the mean ± SD of three independent experiments. P-values vs 0 h: P1 = 4.2 × 10−5, P2 = 7.1 × 10−7, P3 = 6.3 × 10−5, P4 = 2.7 × 10−4, P5 = 6.3 × 10−3, P6 = 2.3 × 10−5, P7 = 1.8 × 10−4, and P8 = 0.044. h EdU incorporation assay to assess global nascent DNA transcription. Representative results of three biological replicates are shown. Blue, <t>DAPI;</t> green, EdU. Scale bar = 20 µm. i CX-5461 induced DNA damage (γ-H2AX, red) was enriched in cells with active DNA synthesis (positive for EdU (green) labeling). Representative results of three biological replicates are shown. Scale bar = 10 μm. Two-tailed t- test was used for (b, c, g). Source data for these panels are included in Source Data file.
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KEYENCE prolong gold antifade reagent dapi
Fig. 3 CX-5461 causes DNA damage and cell death by apoptosis in neuroblastoma cells at sub-micromolar concentrations. a Apoptosis in CHP-134 cells. The cells were treated for 48 h. b Quantification of data in (a). Data represent the mean ± SD of three independent experiments. Significant P-values (left to right): 3.9 × 10−6, 7.2 × 10−7, and 4.8 × 10−7. c Quantification of flow cytometry data showing apoptosis up to 48 h in CHP-134 and IMR-5 (0.2 µM and 0.1 µM of CX-5461 respectively). CHP-134 cells were treated with 0.2 µM and IMR-5 cells with 0.1 µM of CX-5461. Data represent the mean ± SD of three independent experiments. Significant P-values (left to right): CHP-134, 2.2 × 10−4, 3.2 × 10−5, and 1.5 × 10−5; IMR-5, 1.8 × 10−4, 2.3 × 10−5, and 2.5 × 10−6. d Western blots for cleaved PARP (c-PARP). β-Actin was used as a loading control. Representative results of three biological replicates are shown. e Co-localization of 53BP1 and γ-H2AX. Cells were treated for 24 h. Representative results of three biological replicates are shown. Scale bar = 10 μm. See Fig. S3d for more time points. f Comet assay in CHP-134 cells. Tail length (left) and representative images (right) are shown. The red line indicates the mean value of tail length in each group. Scale bar = 5 µm. g Cell cycle analysis forCHP-134 (0.2 µM CX-5461) and IMR-5 (0.05 µM CX-5461) cells. Data represent the mean ± SD of three independent experiments. P-values vs 0 h: P1 = 4.2 × 10−5, P2 = 7.1 × 10−7, P3 = 6.3 × 10−5, P4 = 2.7 × 10−4, P5 = 6.3 × 10−3, P6 = 2.3 × 10−5, P7 = 1.8 × 10−4, and P8 = 0.044. h EdU incorporation assay to assess global nascent DNA transcription. Representative results of three biological replicates are shown. Blue, <t>DAPI;</t> green, EdU. Scale bar = 20 µm. i CX-5461 induced DNA damage (γ-H2AX, red) was enriched in cells with active DNA synthesis (positive for EdU (green) labeling). Representative results of three biological replicates are shown. Scale bar = 10 μm. Two-tailed t- test was used for (b, c, g). Source data for these panels are included in Source Data file.
Prolong Gold Antifade Reagent Dapi, supplied by KEYENCE, 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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Fisher Scientific prolong gold tm anti-fade glass mounting medium dapi
Fig. 3 CX-5461 causes DNA damage and cell death by apoptosis in neuroblastoma cells at sub-micromolar concentrations. a Apoptosis in CHP-134 cells. The cells were treated for 48 h. b Quantification of data in (a). Data represent the mean ± SD of three independent experiments. Significant P-values (left to right): 3.9 × 10−6, 7.2 × 10−7, and 4.8 × 10−7. c Quantification of flow cytometry data showing apoptosis up to 48 h in CHP-134 and IMR-5 (0.2 µM and 0.1 µM of CX-5461 respectively). CHP-134 cells were treated with 0.2 µM and IMR-5 cells with 0.1 µM of CX-5461. Data represent the mean ± SD of three independent experiments. Significant P-values (left to right): CHP-134, 2.2 × 10−4, 3.2 × 10−5, and 1.5 × 10−5; IMR-5, 1.8 × 10−4, 2.3 × 10−5, and 2.5 × 10−6. d Western blots for cleaved PARP (c-PARP). β-Actin was used as a loading control. Representative results of three biological replicates are shown. e Co-localization of 53BP1 and γ-H2AX. Cells were treated for 24 h. Representative results of three biological replicates are shown. Scale bar = 10 μm. See Fig. S3d for more time points. f Comet assay in CHP-134 cells. Tail length (left) and representative images (right) are shown. The red line indicates the mean value of tail length in each group. Scale bar = 5 µm. g Cell cycle analysis forCHP-134 (0.2 µM CX-5461) and IMR-5 (0.05 µM CX-5461) cells. Data represent the mean ± SD of three independent experiments. P-values vs 0 h: P1 = 4.2 × 10−5, P2 = 7.1 × 10−7, P3 = 6.3 × 10−5, P4 = 2.7 × 10−4, P5 = 6.3 × 10−3, P6 = 2.3 × 10−5, P7 = 1.8 × 10−4, and P8 = 0.044. h EdU incorporation assay to assess global nascent DNA transcription. Representative results of three biological replicates are shown. Blue, <t>DAPI;</t> green, EdU. Scale bar = 20 µm. i CX-5461 induced DNA damage (γ-H2AX, red) was enriched in cells with active DNA synthesis (positive for EdU (green) labeling). Representative results of three biological replicates are shown. Scale bar = 10 μm. Two-tailed t- test was used for (b, c, g). Source data for these panels are included in Source Data file.
Prolong Gold Tm Anti Fade Glass Mounting Medium Dapi, supplied by Fisher Scientific, 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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Thermon Group Holdings prolong gold antifade reagent with dapi
a OXPHOS metabolism, reflected by oxygen consumption rate (OCR) levels were measured in MiaPaCa-2 cells after 24 h treatments. b MiaPaCa-2 cells were treated with ZZW-115 (1.5 µM) and olaparib (25 µM) for 24 h, then, loaded with MitoTracker Deep-Red FM and, after fixation, stained with <t>DAPI.</t> Flow cytometry analysis were carried out using MitoProbe™ TMRM ( c ), MitoSOX™ Red ( d ), CellROX™ Orange Reagent ( e ) or Fluo-4-AM ( f ) for analysis of the mitochondrial membrane potential, mitochondrial superoxide levels, intracellular ROS levels and the cytosolic calcium concentration, respectively, after 24 h of incubation with the drugs. Data represent mean ± SEM. Two-way, Sidak was used, * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.001. g PLA was performed in MiaPaCa-2 cells in the presence or in the absence of ZZW-115 together or not with 5-FU in the presence or absence of olaparib. Mouse anti-PAR and rabbit anti-Mitofusin2 antibodies were used. A representative experiment is shown ( n = 3). ImageJ was used to count the number of green dots. Mean ± SEM of foci/nucleus are included.
Prolong Gold Antifade Reagent With Dapi, supplied by Thermon Group Holdings, 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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ZSGB Biotech prolong gold antifade reagent containing 4',6-diamidino-2-phenylindole (dapi) dye
a OXPHOS metabolism, reflected by oxygen consumption rate (OCR) levels were measured in MiaPaCa-2 cells after 24 h treatments. b MiaPaCa-2 cells were treated with ZZW-115 (1.5 µM) and olaparib (25 µM) for 24 h, then, loaded with MitoTracker Deep-Red FM and, after fixation, stained with <t>DAPI.</t> Flow cytometry analysis were carried out using MitoProbe™ TMRM ( c ), MitoSOX™ Red ( d ), CellROX™ Orange Reagent ( e ) or Fluo-4-AM ( f ) for analysis of the mitochondrial membrane potential, mitochondrial superoxide levels, intracellular ROS levels and the cytosolic calcium concentration, respectively, after 24 h of incubation with the drugs. Data represent mean ± SEM. Two-way, Sidak was used, * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.001. g PLA was performed in MiaPaCa-2 cells in the presence or in the absence of ZZW-115 together or not with 5-FU in the presence or absence of olaparib. Mouse anti-PAR and rabbit anti-Mitofusin2 antibodies were used. A representative experiment is shown ( n = 3). ImageJ was used to count the number of green dots. Mean ± SEM of foci/nucleus are included.
Prolong Gold Antifade Reagent Containing 4',6 Diamidino 2 Phenylindole (Dapi) Dye, supplied by ZSGB 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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a OXPHOS metabolism, reflected by oxygen consumption rate (OCR) levels were measured in MiaPaCa-2 cells after 24 h treatments. b MiaPaCa-2 cells were treated with ZZW-115 (1.5 µM) and olaparib (25 µM) for 24 h, then, loaded with MitoTracker Deep-Red FM and, after fixation, stained with <t>DAPI.</t> Flow cytometry analysis were carried out using MitoProbe™ TMRM ( c ), MitoSOX™ Red ( d ), CellROX™ Orange Reagent ( e ) or Fluo-4-AM ( f ) for analysis of the mitochondrial membrane potential, mitochondrial superoxide levels, intracellular ROS levels and the cytosolic calcium concentration, respectively, after 24 h of incubation with the drugs. Data represent mean ± SEM. Two-way, Sidak was used, * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.001. g PLA was performed in MiaPaCa-2 cells in the presence or in the absence of ZZW-115 together or not with 5-FU in the presence or absence of olaparib. Mouse anti-PAR and rabbit anti-Mitofusin2 antibodies were used. A representative experiment is shown ( n = 3). ImageJ was used to count the number of green dots. Mean ± SEM of foci/nucleus are included.
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Keygen Biotech prolong® gold antifade with dapi
a OXPHOS metabolism, reflected by oxygen consumption rate (OCR) levels were measured in MiaPaCa-2 cells after 24 h treatments. b MiaPaCa-2 cells were treated with ZZW-115 (1.5 µM) and olaparib (25 µM) for 24 h, then, loaded with MitoTracker Deep-Red FM and, after fixation, stained with <t>DAPI.</t> Flow cytometry analysis were carried out using MitoProbe™ TMRM ( c ), MitoSOX™ Red ( d ), CellROX™ Orange Reagent ( e ) or Fluo-4-AM ( f ) for analysis of the mitochondrial membrane potential, mitochondrial superoxide levels, intracellular ROS levels and the cytosolic calcium concentration, respectively, after 24 h of incubation with the drugs. Data represent mean ± SEM. Two-way, Sidak was used, * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.001. g PLA was performed in MiaPaCa-2 cells in the presence or in the absence of ZZW-115 together or not with 5-FU in the presence or absence of olaparib. Mouse anti-PAR and rabbit anti-Mitofusin2 antibodies were used. A representative experiment is shown ( n = 3). ImageJ was used to count the number of green dots. Mean ± SEM of foci/nucleus are included.
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a OXPHOS metabolism, reflected by oxygen consumption rate (OCR) levels were measured in MiaPaCa-2 cells after 24 h treatments. b MiaPaCa-2 cells were treated with ZZW-115 (1.5 µM) and olaparib (25 µM) for 24 h, then, loaded with MitoTracker Deep-Red FM and, after fixation, stained with <t>DAPI.</t> Flow cytometry analysis were carried out using MitoProbe™ TMRM ( c ), MitoSOX™ Red ( d ), CellROX™ Orange Reagent ( e ) or Fluo-4-AM ( f ) for analysis of the mitochondrial membrane potential, mitochondrial superoxide levels, intracellular ROS levels and the cytosolic calcium concentration, respectively, after 24 h of incubation with the drugs. Data represent mean ± SEM. Two-way, Sidak was used, * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.001. g PLA was performed in MiaPaCa-2 cells in the presence or in the absence of ZZW-115 together or not with 5-FU in the presence or absence of olaparib. Mouse anti-PAR and rabbit anti-Mitofusin2 antibodies were used. A representative experiment is shown ( n = 3). ImageJ was used to count the number of green dots. Mean ± SEM of foci/nucleus are included.
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Fig. 3 CX-5461 causes DNA damage and cell death by apoptosis in neuroblastoma cells at sub-micromolar concentrations. a Apoptosis in CHP-134 cells. The cells were treated for 48 h. b Quantification of data in (a). Data represent the mean ± SD of three independent experiments. Significant P-values (left to right): 3.9 × 10−6, 7.2 × 10−7, and 4.8 × 10−7. c Quantification of flow cytometry data showing apoptosis up to 48 h in CHP-134 and IMR-5 (0.2 µM and 0.1 µM of CX-5461 respectively). CHP-134 cells were treated with 0.2 µM and IMR-5 cells with 0.1 µM of CX-5461. Data represent the mean ± SD of three independent experiments. Significant P-values (left to right): CHP-134, 2.2 × 10−4, 3.2 × 10−5, and 1.5 × 10−5; IMR-5, 1.8 × 10−4, 2.3 × 10−5, and 2.5 × 10−6. d Western blots for cleaved PARP (c-PARP). β-Actin was used as a loading control. Representative results of three biological replicates are shown. e Co-localization of 53BP1 and γ-H2AX. Cells were treated for 24 h. Representative results of three biological replicates are shown. Scale bar = 10 μm. See Fig. S3d for more time points. f Comet assay in CHP-134 cells. Tail length (left) and representative images (right) are shown. The red line indicates the mean value of tail length in each group. Scale bar = 5 µm. g Cell cycle analysis forCHP-134 (0.2 µM CX-5461) and IMR-5 (0.05 µM CX-5461) cells. Data represent the mean ± SD of three independent experiments. P-values vs 0 h: P1 = 4.2 × 10−5, P2 = 7.1 × 10−7, P3 = 6.3 × 10−5, P4 = 2.7 × 10−4, P5 = 6.3 × 10−3, P6 = 2.3 × 10−5, P7 = 1.8 × 10−4, and P8 = 0.044. h EdU incorporation assay to assess global nascent DNA transcription. Representative results of three biological replicates are shown. Blue, DAPI; green, EdU. Scale bar = 20 µm. i CX-5461 induced DNA damage (γ-H2AX, red) was enriched in cells with active DNA synthesis (positive for EdU (green) labeling). Representative results of three biological replicates are shown. Scale bar = 10 μm. Two-tailed t- test was used for (b, c, g). Source data for these panels are included in Source Data file.

Journal: Nature communications

Article Title: The chemotherapeutic CX-5461 primarily targets TOP2B and exhibits selective activity in high-risk neuroblastoma.

doi: 10.1038/s41467-021-26640-x

Figure Lengend Snippet: Fig. 3 CX-5461 causes DNA damage and cell death by apoptosis in neuroblastoma cells at sub-micromolar concentrations. a Apoptosis in CHP-134 cells. The cells were treated for 48 h. b Quantification of data in (a). Data represent the mean ± SD of three independent experiments. Significant P-values (left to right): 3.9 × 10−6, 7.2 × 10−7, and 4.8 × 10−7. c Quantification of flow cytometry data showing apoptosis up to 48 h in CHP-134 and IMR-5 (0.2 µM and 0.1 µM of CX-5461 respectively). CHP-134 cells were treated with 0.2 µM and IMR-5 cells with 0.1 µM of CX-5461. Data represent the mean ± SD of three independent experiments. Significant P-values (left to right): CHP-134, 2.2 × 10−4, 3.2 × 10−5, and 1.5 × 10−5; IMR-5, 1.8 × 10−4, 2.3 × 10−5, and 2.5 × 10−6. d Western blots for cleaved PARP (c-PARP). β-Actin was used as a loading control. Representative results of three biological replicates are shown. e Co-localization of 53BP1 and γ-H2AX. Cells were treated for 24 h. Representative results of three biological replicates are shown. Scale bar = 10 μm. See Fig. S3d for more time points. f Comet assay in CHP-134 cells. Tail length (left) and representative images (right) are shown. The red line indicates the mean value of tail length in each group. Scale bar = 5 µm. g Cell cycle analysis forCHP-134 (0.2 µM CX-5461) and IMR-5 (0.05 µM CX-5461) cells. Data represent the mean ± SD of three independent experiments. P-values vs 0 h: P1 = 4.2 × 10−5, P2 = 7.1 × 10−7, P3 = 6.3 × 10−5, P4 = 2.7 × 10−4, P5 = 6.3 × 10−3, P6 = 2.3 × 10−5, P7 = 1.8 × 10−4, and P8 = 0.044. h EdU incorporation assay to assess global nascent DNA transcription. Representative results of three biological replicates are shown. Blue, DAPI; green, EdU. Scale bar = 20 µm. i CX-5461 induced DNA damage (γ-H2AX, red) was enriched in cells with active DNA synthesis (positive for EdU (green) labeling). Representative results of three biological replicates are shown. Scale bar = 10 μm. Two-tailed t- test was used for (b, c, g). Source data for these panels are included in Source Data file.

Article Snippet: ProLong® Gold Antifade Reagent with DAPI (Cell Signaling, #8961) was used as coverslip mountant.

Techniques: Cytometry, Western Blot, Control, Single Cell Gel Electrophoresis, Cell Cycle Assay, DNA Synthesis, Labeling, Two Tailed Test

a OXPHOS metabolism, reflected by oxygen consumption rate (OCR) levels were measured in MiaPaCa-2 cells after 24 h treatments. b MiaPaCa-2 cells were treated with ZZW-115 (1.5 µM) and olaparib (25 µM) for 24 h, then, loaded with MitoTracker Deep-Red FM and, after fixation, stained with DAPI. Flow cytometry analysis were carried out using MitoProbe™ TMRM ( c ), MitoSOX™ Red ( d ), CellROX™ Orange Reagent ( e ) or Fluo-4-AM ( f ) for analysis of the mitochondrial membrane potential, mitochondrial superoxide levels, intracellular ROS levels and the cytosolic calcium concentration, respectively, after 24 h of incubation with the drugs. Data represent mean ± SEM. Two-way, Sidak was used, * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.001. g PLA was performed in MiaPaCa-2 cells in the presence or in the absence of ZZW-115 together or not with 5-FU in the presence or absence of olaparib. Mouse anti-PAR and rabbit anti-Mitofusin2 antibodies were used. A representative experiment is shown ( n = 3). ImageJ was used to count the number of green dots. Mean ± SEM of foci/nucleus are included.

Journal: Communications Biology

Article Title: NUPR1 protects against hyperPARylation-dependent cell death

doi: 10.1038/s42003-022-03705-1

Figure Lengend Snippet: a OXPHOS metabolism, reflected by oxygen consumption rate (OCR) levels were measured in MiaPaCa-2 cells after 24 h treatments. b MiaPaCa-2 cells were treated with ZZW-115 (1.5 µM) and olaparib (25 µM) for 24 h, then, loaded with MitoTracker Deep-Red FM and, after fixation, stained with DAPI. Flow cytometry analysis were carried out using MitoProbe™ TMRM ( c ), MitoSOX™ Red ( d ), CellROX™ Orange Reagent ( e ) or Fluo-4-AM ( f ) for analysis of the mitochondrial membrane potential, mitochondrial superoxide levels, intracellular ROS levels and the cytosolic calcium concentration, respectively, after 24 h of incubation with the drugs. Data represent mean ± SEM. Two-way, Sidak was used, * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.001. g PLA was performed in MiaPaCa-2 cells in the presence or in the absence of ZZW-115 together or not with 5-FU in the presence or absence of olaparib. Mouse anti-PAR and rabbit anti-Mitofusin2 antibodies were used. A representative experiment is shown ( n = 3). ImageJ was used to count the number of green dots. Mean ± SEM of foci/nucleus are included.

Article Snippet: Finally, samples were mounted using the Prolong Gold antifade reagent with DAPI (Thermon Fisher).

Techniques: Staining, Flow Cytometry, Membrane, Concentration Assay, Incubation