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dapi  (Miltenyi Biotec)


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    Structured Review

    Miltenyi Biotec dapi
    Dapi, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 97/100, based on 94 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/staining+solution/DAPI+Staining+Solution/pm42855519-284-0-2
    Average 97 stars, based on 94 article reviews
    dapi - by Bioz Stars, 2026-10
    97/100 stars

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    Related Articles

    Flow Cytometry:

    Article Title: Efficient generation of osteoclasts from human induced pluripotent stem cells and functional investigations of lethal CLCN7-related osteopetrosis.
    Article Snippet: .. Flow cytometry Surface marker staining of 5 105 freshly isolated monocytes or harvested hiPSC-derived monocyte-like cells was performed in 50 μl staining solution for 10 min at 4 C. The staining solution contained Miltenyi Biotec (Bergisch Gladbach, Germany) antibodies and reagents diluted as indicated in fluorescenceactivated cell sorting (FACS) buffer (2mM EDTA and 0.5% bovine serum albumin [BSA] in Dulbecco’s phosphate-buffered saline [DPBS]): human Tandem Signal Enhancer 1:17, human Fc receptor [FcR]-Blocking Reagent 1:5, and Isotype controls 1:10 or human surface marker antibodies (CD34-fluorescein isothiocyanate [FITC] 1:10, CD45-VioBlue 1:20, CD66b-phycoerythrin [PE]Vio770 1:20, CD11b-PE 1:50, CD14-allophycocyanin [APC] 1:50). ..

    Article Title: A mechanical non-enzymatic method for isolation of mouse embryonic fibroblasts.
    Article Snippet: Mouse embryonic fibroblasts (MEFs) accessibility coupled with their simple generation make them as a typical embryonic cell model and feeder layer for in vitro expansion of pluripotent stem cells (PSCs).. In this study, a mechanical isolation technique was adopted to isolate MEFs and the efficiency of this technique was compared with enzymatic digestion method.. The suspended MEFs were prepared either by mechanical method or 0.25% trypsin enzymatic digestion.

    Marker:

    Article Title: Efficient generation of osteoclasts from human induced pluripotent stem cells and functional investigations of lethal CLCN7-related osteopetrosis.
    Article Snippet: .. Flow cytometry Surface marker staining of 5 105 freshly isolated monocytes or harvested hiPSC-derived monocyte-like cells was performed in 50 μl staining solution for 10 min at 4 C. The staining solution contained Miltenyi Biotec (Bergisch Gladbach, Germany) antibodies and reagents diluted as indicated in fluorescenceactivated cell sorting (FACS) buffer (2mM EDTA and 0.5% bovine serum albumin [BSA] in Dulbecco’s phosphate-buffered saline [DPBS]): human Tandem Signal Enhancer 1:17, human Fc receptor [FcR]-Blocking Reagent 1:5, and Isotype controls 1:10 or human surface marker antibodies (CD34-fluorescein isothiocyanate [FITC] 1:10, CD45-VioBlue 1:20, CD66b-phycoerythrin [PE]Vio770 1:20, CD11b-PE 1:50, CD14-allophycocyanin [APC] 1:50). ..

    Article Title: Focused ultrasound-induced blood-brain barrier opening promotes glioprotective phenotypes in ACSA-II+ murine astrocytes
    Article Snippet: .. First, the staining solution consisted of ACSA-II-PE astrocytic marker (Miltenyi #130-123-284) and anti-myelin oligodendrocyte glycoprotein (MOG) AF488 (Abcam, #ab306602) to evaluate the astrocyte and mature, myelinating oligodendrocyte populations simultaneously. ..

    Staining:

    Article Title: Efficient generation of osteoclasts from human induced pluripotent stem cells and functional investigations of lethal CLCN7-related osteopetrosis.
    Article Snippet: .. Flow cytometry Surface marker staining of 5 105 freshly isolated monocytes or harvested hiPSC-derived monocyte-like cells was performed in 50 μl staining solution for 10 min at 4 C. The staining solution contained Miltenyi Biotec (Bergisch Gladbach, Germany) antibodies and reagents diluted as indicated in fluorescenceactivated cell sorting (FACS) buffer (2mM EDTA and 0.5% bovine serum albumin [BSA] in Dulbecco’s phosphate-buffered saline [DPBS]): human Tandem Signal Enhancer 1:17, human Fc receptor [FcR]-Blocking Reagent 1:5, and Isotype controls 1:10 or human surface marker antibodies (CD34-fluorescein isothiocyanate [FITC] 1:10, CD45-VioBlue 1:20, CD66b-phycoerythrin [PE]Vio770 1:20, CD11b-PE 1:50, CD14-allophycocyanin [APC] 1:50). ..

    Article Title: Focused ultrasound-induced blood-brain barrier opening promotes glioprotective phenotypes in ACSA-II+ murine astrocytes
    Article Snippet: .. First, the staining solution consisted of ACSA-II-PE astrocytic marker (Miltenyi #130-123-284) and anti-myelin oligodendrocyte glycoprotein (MOG) AF488 (Abcam, #ab306602) to evaluate the astrocyte and mature, myelinating oligodendrocyte populations simultaneously. ..

    Article Title: Doxorubicin treatment modulates chemoresistance and affects the cell cycle in two canine mammary tumour cell lines
    Article Snippet: Aliquots of 1 × 10 6 cells were washed twice in 5 ml of PBS w/o Ca 2+ and Mg 2+ (Gibco-Life Technologies) and fixed overnight in 70% ice-cold ethanol (1 ml) added drop-by-drop with continuous vortexing. .. Then, the cells were washed with 10 ml of PBS and cellular pellet was incubated with 1 ml of staining solution containing 50 μg/ml of PI (Miltenyi Biotec, Bergisch Gladbach, Germany) and 100 μg/ml RNaseA/T1 (Thermo Scientific) in PBS for 30 min in the dark at room temperature (RT). .. Cell distribution in cell cycle phases was analyzed by MACSQuant® Analyzer10 and Flowlogic software (Miltenyi Biotec).

    Article Title: Comprehensive Evaluation of Biological Effects of Pentathiepins on Various Human Cancer Cell Lines and Insights into Their Mode of Action
    Article Snippet: .. The staining solution was incubated for 30 min, and the cell suspension was measured at λ Ex / Em = 488/655–730 nm using a MACSquant Analyzer 10 (Miltenyi Biotec, Teterow, Germany). .. All Western blot analyses were performed with reagents, material, and devices purchased from Bio-Rad (Feldkirchen, Germany) if not stated otherwise.

    Article Title: Berberine Inhibits Telomerase Activity and Induces Cell Cycle Arrest and Telomere Erosion in Colorectal Cancer Cell Line, HCT 116
    Article Snippet: .. The suspension was stored at 4 °C for 24 h. After fixation, 1 mL of sample buffer was added while vortexing, followed by centrifugation at 17,000× g for 10 min. Staining solution made up of 0.5 μg/mL 4′,6 -diamidino-2-phenylindole (DAPI) (Miltenyi Biotec, Bergisch Gladbach, Germany), 5 μg/mL RNase A, and 0.1% Tween 20 (Classic Chemicals, Shah Alam, Selangor, Malaysia) in 1× PBS was prepared immediately before incubation. .. The supernatant was removed, and 1 mL of staining solution was added followed by vortexing and incubation at room temperature for 40 min. Flow cytometry was done by using a MACSQuant ® Analyzer (Miltenyi Biotec, Bergisch Gladbach, Germany) following the protocol suggested by Miltenyi Biotec with optimizations.

    Article Title: Transcriptional CDK Inhibitors CYC065 and THZ1 Induce Apoptosis in Glioma Stem Cells Derived from Recurrent GBM
    Article Snippet: GBM cells were cultured as gliomaspheres and left untreated for basal expression analysis or treated with 150 μM TMZ (#T2577, Sigma-Aldrich, Arklow, Ireland), 3 μM CYC065 (#HY-101212, Medchemexpress, NJ, USA) or 100 nM THZ1 (#HY-80013, Medchemexpress, NJ, USA) for 120 h. At the end of treatment window, cells were harvested, dissociated using Accutase (Thermo Fisher Scientific, Waltham, MA, USA) and washed in FACS washing media (DPBS + 5% FCS, Gibco Life Technologies, Dún Laoghaire, Ireland). .. Cells were incubated for 15 min in FACS wash media on ice and stained with staining solution: unstained control, CD133-VioBright 667 (#130-111-756, MACS, Miltenyi Biotec, Bisley, UK), VioBright 667-isotype control (#130-118-217, MACS, Miltenyi Biotec, Bisley, UK), CD44-FITC (#555478, BD Pharmingen, Franklin Lakes, NJ, USA), FITC-isotype control (#555748, BD Pharmingen, Franklin Lakes, NJ, USA) for 30 min on ice, protected from light. ..

    Article Title: Triple Selection Strategy for In Situ Labeling of Circulating Tumor Cells with High Purity and Viability toward Preclinical Personalized Drug Sensitivity Analysis.
    Article Snippet: DOI: 10.1002/adbi.202000013 clinical samples for analysis.. For example, identification and quantification of circulating tumor cells (CTCs)[1–3] or circulating tumor DNA (ctDNA)[4–6] from the blood samples have become complementary biomarkers to aid the selection of suitable treatment for patients.. [7] The enumeration analysis and molecular profiling of these extremely rare CTCs from patient blood samples (1–10 CTCs out of 109 blood cells mL−1) have been applied to the prostate, lung, breast, colon and other types of cancer.

    Article Title: A mechanical non-enzymatic method for isolation of mouse embryonic fibroblasts.
    Article Snippet: Mouse embryonic fibroblasts (MEFs) accessibility coupled with their simple generation make them as a typical embryonic cell model and feeder layer for in vitro expansion of pluripotent stem cells (PSCs).. In this study, a mechanical isolation technique was adopted to isolate MEFs and the efficiency of this technique was compared with enzymatic digestion method.. The suspended MEFs were prepared either by mechanical method or 0.25% trypsin enzymatic digestion.

    Isolation:

    Article Title: Efficient generation of osteoclasts from human induced pluripotent stem cells and functional investigations of lethal CLCN7-related osteopetrosis.
    Article Snippet: .. Flow cytometry Surface marker staining of 5 105 freshly isolated monocytes or harvested hiPSC-derived monocyte-like cells was performed in 50 μl staining solution for 10 min at 4 C. The staining solution contained Miltenyi Biotec (Bergisch Gladbach, Germany) antibodies and reagents diluted as indicated in fluorescenceactivated cell sorting (FACS) buffer (2mM EDTA and 0.5% bovine serum albumin [BSA] in Dulbecco’s phosphate-buffered saline [DPBS]): human Tandem Signal Enhancer 1:17, human Fc receptor [FcR]-Blocking Reagent 1:5, and Isotype controls 1:10 or human surface marker antibodies (CD34-fluorescein isothiocyanate [FITC] 1:10, CD45-VioBlue 1:20, CD66b-phycoerythrin [PE]Vio770 1:20, CD11b-PE 1:50, CD14-allophycocyanin [APC] 1:50). ..

    FACS:

    Article Title: Efficient generation of osteoclasts from human induced pluripotent stem cells and functional investigations of lethal CLCN7-related osteopetrosis.
    Article Snippet: .. Flow cytometry Surface marker staining of 5 105 freshly isolated monocytes or harvested hiPSC-derived monocyte-like cells was performed in 50 μl staining solution for 10 min at 4 C. The staining solution contained Miltenyi Biotec (Bergisch Gladbach, Germany) antibodies and reagents diluted as indicated in fluorescenceactivated cell sorting (FACS) buffer (2mM EDTA and 0.5% bovine serum albumin [BSA] in Dulbecco’s phosphate-buffered saline [DPBS]): human Tandem Signal Enhancer 1:17, human Fc receptor [FcR]-Blocking Reagent 1:5, and Isotype controls 1:10 or human surface marker antibodies (CD34-fluorescein isothiocyanate [FITC] 1:10, CD45-VioBlue 1:20, CD66b-phycoerythrin [PE]Vio770 1:20, CD11b-PE 1:50, CD14-allophycocyanin [APC] 1:50). ..

    Article Title: Transcriptional CDK Inhibitors CYC065 and THZ1 Induce Apoptosis in Glioma Stem Cells Derived from Recurrent GBM
    Article Snippet: GBM cells were cultured as gliomaspheres and left untreated for basal expression analysis or treated with 150 μM TMZ (#T2577, Sigma-Aldrich, Arklow, Ireland), 3 μM CYC065 (#HY-101212, Medchemexpress, NJ, USA) or 100 nM THZ1 (#HY-80013, Medchemexpress, NJ, USA) for 120 h. At the end of treatment window, cells were harvested, dissociated using Accutase (Thermo Fisher Scientific, Waltham, MA, USA) and washed in FACS washing media (DPBS + 5% FCS, Gibco Life Technologies, Dún Laoghaire, Ireland). .. Cells were incubated for 15 min in FACS wash media on ice and stained with staining solution: unstained control, CD133-VioBright 667 (#130-111-756, MACS, Miltenyi Biotec, Bisley, UK), VioBright 667-isotype control (#130-118-217, MACS, Miltenyi Biotec, Bisley, UK), CD44-FITC (#555478, BD Pharmingen, Franklin Lakes, NJ, USA), FITC-isotype control (#555748, BD Pharmingen, Franklin Lakes, NJ, USA) for 30 min on ice, protected from light. ..

    Saline:

    Article Title: Efficient generation of osteoclasts from human induced pluripotent stem cells and functional investigations of lethal CLCN7-related osteopetrosis.
    Article Snippet: .. Flow cytometry Surface marker staining of 5 105 freshly isolated monocytes or harvested hiPSC-derived monocyte-like cells was performed in 50 μl staining solution for 10 min at 4 C. The staining solution contained Miltenyi Biotec (Bergisch Gladbach, Germany) antibodies and reagents diluted as indicated in fluorescenceactivated cell sorting (FACS) buffer (2mM EDTA and 0.5% bovine serum albumin [BSA] in Dulbecco’s phosphate-buffered saline [DPBS]): human Tandem Signal Enhancer 1:17, human Fc receptor [FcR]-Blocking Reagent 1:5, and Isotype controls 1:10 or human surface marker antibodies (CD34-fluorescein isothiocyanate [FITC] 1:10, CD45-VioBlue 1:20, CD66b-phycoerythrin [PE]Vio770 1:20, CD11b-PE 1:50, CD14-allophycocyanin [APC] 1:50). ..

    Incubation:

    Article Title: Doxorubicin treatment modulates chemoresistance and affects the cell cycle in two canine mammary tumour cell lines
    Article Snippet: Aliquots of 1 × 10 6 cells were washed twice in 5 ml of PBS w/o Ca 2+ and Mg 2+ (Gibco-Life Technologies) and fixed overnight in 70% ice-cold ethanol (1 ml) added drop-by-drop with continuous vortexing. .. Then, the cells were washed with 10 ml of PBS and cellular pellet was incubated with 1 ml of staining solution containing 50 μg/ml of PI (Miltenyi Biotec, Bergisch Gladbach, Germany) and 100 μg/ml RNaseA/T1 (Thermo Scientific) in PBS for 30 min in the dark at room temperature (RT). .. Cell distribution in cell cycle phases was analyzed by MACSQuant® Analyzer10 and Flowlogic software (Miltenyi Biotec).

    Article Title: Comprehensive Evaluation of Biological Effects of Pentathiepins on Various Human Cancer Cell Lines and Insights into Their Mode of Action
    Article Snippet: .. The staining solution was incubated for 30 min, and the cell suspension was measured at λ Ex / Em = 488/655–730 nm using a MACSquant Analyzer 10 (Miltenyi Biotec, Teterow, Germany). .. All Western blot analyses were performed with reagents, material, and devices purchased from Bio-Rad (Feldkirchen, Germany) if not stated otherwise.

    Article Title: Berberine Inhibits Telomerase Activity and Induces Cell Cycle Arrest and Telomere Erosion in Colorectal Cancer Cell Line, HCT 116
    Article Snippet: .. The suspension was stored at 4 °C for 24 h. After fixation, 1 mL of sample buffer was added while vortexing, followed by centrifugation at 17,000× g for 10 min. Staining solution made up of 0.5 μg/mL 4′,6 -diamidino-2-phenylindole (DAPI) (Miltenyi Biotec, Bergisch Gladbach, Germany), 5 μg/mL RNase A, and 0.1% Tween 20 (Classic Chemicals, Shah Alam, Selangor, Malaysia) in 1× PBS was prepared immediately before incubation. .. The supernatant was removed, and 1 mL of staining solution was added followed by vortexing and incubation at room temperature for 40 min. Flow cytometry was done by using a MACSQuant ® Analyzer (Miltenyi Biotec, Bergisch Gladbach, Germany) following the protocol suggested by Miltenyi Biotec with optimizations.

    Article Title: Transcriptional CDK Inhibitors CYC065 and THZ1 Induce Apoptosis in Glioma Stem Cells Derived from Recurrent GBM
    Article Snippet: GBM cells were cultured as gliomaspheres and left untreated for basal expression analysis or treated with 150 μM TMZ (#T2577, Sigma-Aldrich, Arklow, Ireland), 3 μM CYC065 (#HY-101212, Medchemexpress, NJ, USA) or 100 nM THZ1 (#HY-80013, Medchemexpress, NJ, USA) for 120 h. At the end of treatment window, cells were harvested, dissociated using Accutase (Thermo Fisher Scientific, Waltham, MA, USA) and washed in FACS washing media (DPBS + 5% FCS, Gibco Life Technologies, Dún Laoghaire, Ireland). .. Cells were incubated for 15 min in FACS wash media on ice and stained with staining solution: unstained control, CD133-VioBright 667 (#130-111-756, MACS, Miltenyi Biotec, Bisley, UK), VioBright 667-isotype control (#130-118-217, MACS, Miltenyi Biotec, Bisley, UK), CD44-FITC (#555478, BD Pharmingen, Franklin Lakes, NJ, USA), FITC-isotype control (#555748, BD Pharmingen, Franklin Lakes, NJ, USA) for 30 min on ice, protected from light. ..

    Article Title: Triple Selection Strategy for In Situ Labeling of Circulating Tumor Cells with High Purity and Viability toward Preclinical Personalized Drug Sensitivity Analysis.
    Article Snippet: DOI: 10.1002/adbi.202000013 clinical samples for analysis.. For example, identification and quantification of circulating tumor cells (CTCs)[1–3] or circulating tumor DNA (ctDNA)[4–6] from the blood samples have become complementary biomarkers to aid the selection of suitable treatment for patients.. [7] The enumeration analysis and molecular profiling of these extremely rare CTCs from patient blood samples (1–10 CTCs out of 109 blood cells mL−1) have been applied to the prostate, lung, breast, colon and other types of cancer.

    Article Title: A mechanical non-enzymatic method for isolation of mouse embryonic fibroblasts.
    Article Snippet: Mouse embryonic fibroblasts (MEFs) accessibility coupled with their simple generation make them as a typical embryonic cell model and feeder layer for in vitro expansion of pluripotent stem cells (PSCs).. In this study, a mechanical isolation technique was adopted to isolate MEFs and the efficiency of this technique was compared with enzymatic digestion method.. The suspended MEFs were prepared either by mechanical method or 0.25% trypsin enzymatic digestion.

    Suspension:

    Article Title: Comprehensive Evaluation of Biological Effects of Pentathiepins on Various Human Cancer Cell Lines and Insights into Their Mode of Action
    Article Snippet: .. The staining solution was incubated for 30 min, and the cell suspension was measured at λ Ex / Em = 488/655–730 nm using a MACSquant Analyzer 10 (Miltenyi Biotec, Teterow, Germany). .. All Western blot analyses were performed with reagents, material, and devices purchased from Bio-Rad (Feldkirchen, Germany) if not stated otherwise.

    Article Title: Berberine Inhibits Telomerase Activity and Induces Cell Cycle Arrest and Telomere Erosion in Colorectal Cancer Cell Line, HCT 116
    Article Snippet: .. The suspension was stored at 4 °C for 24 h. After fixation, 1 mL of sample buffer was added while vortexing, followed by centrifugation at 17,000× g for 10 min. Staining solution made up of 0.5 μg/mL 4′,6 -diamidino-2-phenylindole (DAPI) (Miltenyi Biotec, Bergisch Gladbach, Germany), 5 μg/mL RNase A, and 0.1% Tween 20 (Classic Chemicals, Shah Alam, Selangor, Malaysia) in 1× PBS was prepared immediately before incubation. .. The supernatant was removed, and 1 mL of staining solution was added followed by vortexing and incubation at room temperature for 40 min. Flow cytometry was done by using a MACSQuant ® Analyzer (Miltenyi Biotec, Bergisch Gladbach, Germany) following the protocol suggested by Miltenyi Biotec with optimizations.

    Centrifugation:

    Article Title: Berberine Inhibits Telomerase Activity and Induces Cell Cycle Arrest and Telomere Erosion in Colorectal Cancer Cell Line, HCT 116
    Article Snippet: .. The suspension was stored at 4 °C for 24 h. After fixation, 1 mL of sample buffer was added while vortexing, followed by centrifugation at 17,000× g for 10 min. Staining solution made up of 0.5 μg/mL 4′,6 -diamidino-2-phenylindole (DAPI) (Miltenyi Biotec, Bergisch Gladbach, Germany), 5 μg/mL RNase A, and 0.1% Tween 20 (Classic Chemicals, Shah Alam, Selangor, Malaysia) in 1× PBS was prepared immediately before incubation. .. The supernatant was removed, and 1 mL of staining solution was added followed by vortexing and incubation at room temperature for 40 min. Flow cytometry was done by using a MACSQuant ® Analyzer (Miltenyi Biotec, Bergisch Gladbach, Germany) following the protocol suggested by Miltenyi Biotec with optimizations.

    Control:

    Article Title: Transcriptional CDK Inhibitors CYC065 and THZ1 Induce Apoptosis in Glioma Stem Cells Derived from Recurrent GBM
    Article Snippet: GBM cells were cultured as gliomaspheres and left untreated for basal expression analysis or treated with 150 μM TMZ (#T2577, Sigma-Aldrich, Arklow, Ireland), 3 μM CYC065 (#HY-101212, Medchemexpress, NJ, USA) or 100 nM THZ1 (#HY-80013, Medchemexpress, NJ, USA) for 120 h. At the end of treatment window, cells were harvested, dissociated using Accutase (Thermo Fisher Scientific, Waltham, MA, USA) and washed in FACS washing media (DPBS + 5% FCS, Gibco Life Technologies, Dún Laoghaire, Ireland). .. Cells were incubated for 15 min in FACS wash media on ice and stained with staining solution: unstained control, CD133-VioBright 667 (#130-111-756, MACS, Miltenyi Biotec, Bisley, UK), VioBright 667-isotype control (#130-118-217, MACS, Miltenyi Biotec, Bisley, UK), CD44-FITC (#555478, BD Pharmingen, Franklin Lakes, NJ, USA), FITC-isotype control (#555748, BD Pharmingen, Franklin Lakes, NJ, USA) for 30 min on ice, protected from light. ..

    Magnetic Cell Separation:

    Article Title: Transcriptional CDK Inhibitors CYC065 and THZ1 Induce Apoptosis in Glioma Stem Cells Derived from Recurrent GBM
    Article Snippet: GBM cells were cultured as gliomaspheres and left untreated for basal expression analysis or treated with 150 μM TMZ (#T2577, Sigma-Aldrich, Arklow, Ireland), 3 μM CYC065 (#HY-101212, Medchemexpress, NJ, USA) or 100 nM THZ1 (#HY-80013, Medchemexpress, NJ, USA) for 120 h. At the end of treatment window, cells were harvested, dissociated using Accutase (Thermo Fisher Scientific, Waltham, MA, USA) and washed in FACS washing media (DPBS + 5% FCS, Gibco Life Technologies, Dún Laoghaire, Ireland). .. Cells were incubated for 15 min in FACS wash media on ice and stained with staining solution: unstained control, CD133-VioBright 667 (#130-111-756, MACS, Miltenyi Biotec, Bisley, UK), VioBright 667-isotype control (#130-118-217, MACS, Miltenyi Biotec, Bisley, UK), CD44-FITC (#555478, BD Pharmingen, Franklin Lakes, NJ, USA), FITC-isotype control (#555748, BD Pharmingen, Franklin Lakes, NJ, USA) for 30 min on ice, protected from light. ..



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    Beyotime alizarin red s staining
    HG and PA treatments induce ROS over-production, oxidative stress, and mitochondrial dysfunction in macrophages. ( A ) Representative intracellular ROS photographs using DCFH-DA fluorescence probe in control RAW264.7 macrophages and those with treatments of 25 mmol/L HG and 60 μM PA for 48 h. Scale bars, 50 μm. ( B ) Quantification of fluorescence intensity of ROS in control and HG- and PA-induced RAW264.7 macrophages. ( C and D ) Measurement of MDA and GSH levels in control and HG- and PA-induced RAW264.7 macrophages. ( E ) <t>Representative</t> <t>JC-1</t> staining photographs in control and HG- and PA-induced RAW264.7 macrophages. Green fluorescence represents that MMP is low and JC-1 exists in the cytoplasm in the form of monomer. Red fluorescence represents that MMP is high and JC-1 polymer is formed. Scale bars, 50 μm. ( F ) Calculation of a ratio of green fluorescence and red fluorescence for quantification MMP changes. ( G ) Detection of ATP in control and HG- and PA-induced RAW264.7 macrophages. ( H ) TEM for investigating the changes in mitochondrial morphology in control and HG- and PA-induced RAW264.7 macrophages. Scale bars, 500 nm. **, P<0.01; ***, P<0.001; ****, P<0.0001 derived from Student’s t test.
    Alizarin Red S Staining, supplied by Beyotime, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    HG and PA treatments induce ROS over-production, oxidative stress, and mitochondrial dysfunction in macrophages. ( A ) Representative intracellular ROS photographs using DCFH-DA fluorescence probe in control RAW264.7 macrophages and those with treatments of 25 mmol/L HG and 60 μM PA for 48 h. Scale bars, 50 μm. ( B ) Quantification of fluorescence intensity of ROS in control and HG- and PA-induced RAW264.7 macrophages. ( C and D ) Measurement of MDA and GSH levels in control and HG- and PA-induced RAW264.7 macrophages. ( E ) Representative JC-1 staining photographs in control and HG- and PA-induced RAW264.7 macrophages. Green fluorescence represents that MMP is low and JC-1 exists in the cytoplasm in the form of monomer. Red fluorescence represents that MMP is high and JC-1 polymer is formed. Scale bars, 50 μm. ( F ) Calculation of a ratio of green fluorescence and red fluorescence for quantification MMP changes. ( G ) Detection of ATP in control and HG- and PA-induced RAW264.7 macrophages. ( H ) TEM for investigating the changes in mitochondrial morphology in control and HG- and PA-induced RAW264.7 macrophages. Scale bars, 500 nm. **, P<0.01; ***, P<0.001; ****, P<0.0001 derived from Student’s t test.

    Journal: Journal of Inflammation Research

    Article Title: HMGCS2 Palmitoylation Promotes Diabetes-Induced Erectile Dysfunction Through PPARα-Mediated Macrophage Extracellular Trap Formation

    doi: 10.2147/JIR.S623377

    Figure Lengend Snippet: HG and PA treatments induce ROS over-production, oxidative stress, and mitochondrial dysfunction in macrophages. ( A ) Representative intracellular ROS photographs using DCFH-DA fluorescence probe in control RAW264.7 macrophages and those with treatments of 25 mmol/L HG and 60 μM PA for 48 h. Scale bars, 50 μm. ( B ) Quantification of fluorescence intensity of ROS in control and HG- and PA-induced RAW264.7 macrophages. ( C and D ) Measurement of MDA and GSH levels in control and HG- and PA-induced RAW264.7 macrophages. ( E ) Representative JC-1 staining photographs in control and HG- and PA-induced RAW264.7 macrophages. Green fluorescence represents that MMP is low and JC-1 exists in the cytoplasm in the form of monomer. Red fluorescence represents that MMP is high and JC-1 polymer is formed. Scale bars, 50 μm. ( F ) Calculation of a ratio of green fluorescence and red fluorescence for quantification MMP changes. ( G ) Detection of ATP in control and HG- and PA-induced RAW264.7 macrophages. ( H ) TEM for investigating the changes in mitochondrial morphology in control and HG- and PA-induced RAW264.7 macrophages. Scale bars, 500 nm. **, P<0.01; ***, P<0.001; ****, P<0.0001 derived from Student’s t test.

    Article Snippet: After 24 h, 300 μL JC-1 staining solution (C2006; Beyotime) was added, with additional 20-min cultivation in the dark at 37°C.

    Techniques: Fluorescence, Control, Staining, Polymer, Morphology, Derivative Assay

    Pharmacological suppression of protein palmitoylation attenuates HG- and PA-induced ROS over-production, oxidative stress, and mitochondrial dysfunction in macrophages. ( A ) Representative intracellular ROS photographs by the use of DCFH-DA fluorescence probe in control macrophages, HG- and PA-induced macrophages, and HG- and PA-induced macrophages with CMA treatment. Scale bars, 50 μm. ( B ) Quantification of fluorescence intensity of ROS in control macrophages, HG- and PA-induced macrophages, and HG- and PA-induced macrophages with CMA treatment. ( C and D ) Measurement of MDA and GSH levels in control macrophages, HG- and PA-induced macrophages, and HG- and PA-induced macrophages with CMA treatment. ( E ) Representative JC-1 staining photographs in control macrophages, HG- and PA-induced macrophages, and HG- and PA-induced macrophages with CMA treatment. Green fluorescence represents that MMP is low and JC-1 exists in the cytoplasm in the form of monomer. Red fluorescence represents that MMP is high and JC-1 polymer is formed. Scale bars, 50 μm. ( F ) Calculation of a ratio of green fluorescence and red fluorescence for quantifying MMP changes. ( G ) Detection of ATP in control macrophages, HG- and PA-induced macrophages, and HG- and PA-induced macrophages with CMA treatment. ( H ) TEM for investigating the changes in mitochondrial morphology in control macrophages, HG- and PA-induced macrophages, and HG- and PA-induced macrophages with CMA treatment. Scale bars, 500 nm. *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001 derived from one-way ANOVA followed by Tukey’s post hoc test.

    Journal: Journal of Inflammation Research

    Article Title: HMGCS2 Palmitoylation Promotes Diabetes-Induced Erectile Dysfunction Through PPARα-Mediated Macrophage Extracellular Trap Formation

    doi: 10.2147/JIR.S623377

    Figure Lengend Snippet: Pharmacological suppression of protein palmitoylation attenuates HG- and PA-induced ROS over-production, oxidative stress, and mitochondrial dysfunction in macrophages. ( A ) Representative intracellular ROS photographs by the use of DCFH-DA fluorescence probe in control macrophages, HG- and PA-induced macrophages, and HG- and PA-induced macrophages with CMA treatment. Scale bars, 50 μm. ( B ) Quantification of fluorescence intensity of ROS in control macrophages, HG- and PA-induced macrophages, and HG- and PA-induced macrophages with CMA treatment. ( C and D ) Measurement of MDA and GSH levels in control macrophages, HG- and PA-induced macrophages, and HG- and PA-induced macrophages with CMA treatment. ( E ) Representative JC-1 staining photographs in control macrophages, HG- and PA-induced macrophages, and HG- and PA-induced macrophages with CMA treatment. Green fluorescence represents that MMP is low and JC-1 exists in the cytoplasm in the form of monomer. Red fluorescence represents that MMP is high and JC-1 polymer is formed. Scale bars, 50 μm. ( F ) Calculation of a ratio of green fluorescence and red fluorescence for quantifying MMP changes. ( G ) Detection of ATP in control macrophages, HG- and PA-induced macrophages, and HG- and PA-induced macrophages with CMA treatment. ( H ) TEM for investigating the changes in mitochondrial morphology in control macrophages, HG- and PA-induced macrophages, and HG- and PA-induced macrophages with CMA treatment. Scale bars, 500 nm. *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001 derived from one-way ANOVA followed by Tukey’s post hoc test.

    Article Snippet: After 24 h, 300 μL JC-1 staining solution (C2006; Beyotime) was added, with additional 20-min cultivation in the dark at 37°C.

    Techniques: Fluorescence, Control, Staining, Polymer, Morphology, Derivative Assay