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Millipore etbr
Increased leakage of mtDNA into the cytosol promoted STING activation in aged macrophages. (a, b) Young and aged BMDMs were subjected to HR as described in the section of methods. (a) Immunoblotting of cGAS, P‐STING S365, P‐TBK1 S172, TBK1, P‐NF‐κB S536, and GAPDH. (b) ELISA analysis of TNF‐α and IL‐6 in medium supernatant ( n = 3). (c) ELISA analysis of intracellular 2,3‐cGAMP ( n = 3). (d) Confocal microscopy images of BMDMs stained with MitoTracker Red and PicoGreen (scale bar, 10 μm). Relative fluorescence intensity of cytosolic mtDNA was plotted ( n = 5). Tunicamycin served as a positive control (4 μg/ml). (e) qRT‐PCR assessment for cytosolic mtDNA ( n = 3). (f–i) The BMDMs were treated with <t>EtBr</t> <t>(400ng/ml).</t> (f) Immunofluorescent imaging (scale bar, 10 μm) and (g) qRT‐PCR analysis of cytosolic mtDNA confirming mtDNA depletion by EtBr ( n = 3). (h) Immunoblotting of cGAS, P‐STING S365, P‐TBK1 S172, TBK1, P‐NF‐κB S536, and GAPDH. (i) ELISA analysis of TNF‐α and IL‐6 in medium supernatant ( n = 3). (j) ELISA analysis of intracellular 2,3‐cGAMP ( n = 3). (k–m) The BMDMs were pre‐transfected with cGAS siRNA or Ns siRNA for 36 h before HR as described in the section of methods. (k) Immunoblotting of cGAS, P‐STING S365, P‐TBK1 S172, TBK1, P‐NF‐κB S536, and GAPDH. (l) ELISA analysis of TNF‐α and IL‐6 in medium supernatant ( n = 3). (m) ELISA analysis of intracellular 2,3‐cGAMP ( n = 3). All these immunoblots have been repeated for 3 times. Data are presented as mean ± SD. * p
Etbr, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Defective mitophagy in aged macrophages promotes mitochondrial DNA cytosolic leakage to activate STING signaling during liver sterile inflammation). Defective mitophagy in aged macrophages promotes mitochondrial DNA cytosolic leakage to activate STING signaling during liver sterile inflammation"

Article Title: Defective mitophagy in aged macrophages promotes mitochondrial DNA cytosolic leakage to activate STING signaling during liver sterile inflammation). Defective mitophagy in aged macrophages promotes mitochondrial DNA cytosolic leakage to activate STING signaling during liver sterile inflammation

Journal: Aging Cell

doi: 10.1111/acel.13622

Increased leakage of mtDNA into the cytosol promoted STING activation in aged macrophages. (a, b) Young and aged BMDMs were subjected to HR as described in the section of methods. (a) Immunoblotting of cGAS, P‐STING S365, P‐TBK1 S172, TBK1, P‐NF‐κB S536, and GAPDH. (b) ELISA analysis of TNF‐α and IL‐6 in medium supernatant ( n = 3). (c) ELISA analysis of intracellular 2,3‐cGAMP ( n = 3). (d) Confocal microscopy images of BMDMs stained with MitoTracker Red and PicoGreen (scale bar, 10 μm). Relative fluorescence intensity of cytosolic mtDNA was plotted ( n = 5). Tunicamycin served as a positive control (4 μg/ml). (e) qRT‐PCR assessment for cytosolic mtDNA ( n = 3). (f–i) The BMDMs were treated with EtBr (400ng/ml). (f) Immunofluorescent imaging (scale bar, 10 μm) and (g) qRT‐PCR analysis of cytosolic mtDNA confirming mtDNA depletion by EtBr ( n = 3). (h) Immunoblotting of cGAS, P‐STING S365, P‐TBK1 S172, TBK1, P‐NF‐κB S536, and GAPDH. (i) ELISA analysis of TNF‐α and IL‐6 in medium supernatant ( n = 3). (j) ELISA analysis of intracellular 2,3‐cGAMP ( n = 3). (k–m) The BMDMs were pre‐transfected with cGAS siRNA or Ns siRNA for 36 h before HR as described in the section of methods. (k) Immunoblotting of cGAS, P‐STING S365, P‐TBK1 S172, TBK1, P‐NF‐κB S536, and GAPDH. (l) ELISA analysis of TNF‐α and IL‐6 in medium supernatant ( n = 3). (m) ELISA analysis of intracellular 2,3‐cGAMP ( n = 3). All these immunoblots have been repeated for 3 times. Data are presented as mean ± SD. * p
Figure Legend Snippet: Increased leakage of mtDNA into the cytosol promoted STING activation in aged macrophages. (a, b) Young and aged BMDMs were subjected to HR as described in the section of methods. (a) Immunoblotting of cGAS, P‐STING S365, P‐TBK1 S172, TBK1, P‐NF‐κB S536, and GAPDH. (b) ELISA analysis of TNF‐α and IL‐6 in medium supernatant ( n = 3). (c) ELISA analysis of intracellular 2,3‐cGAMP ( n = 3). (d) Confocal microscopy images of BMDMs stained with MitoTracker Red and PicoGreen (scale bar, 10 μm). Relative fluorescence intensity of cytosolic mtDNA was plotted ( n = 5). Tunicamycin served as a positive control (4 μg/ml). (e) qRT‐PCR assessment for cytosolic mtDNA ( n = 3). (f–i) The BMDMs were treated with EtBr (400ng/ml). (f) Immunofluorescent imaging (scale bar, 10 μm) and (g) qRT‐PCR analysis of cytosolic mtDNA confirming mtDNA depletion by EtBr ( n = 3). (h) Immunoblotting of cGAS, P‐STING S365, P‐TBK1 S172, TBK1, P‐NF‐κB S536, and GAPDH. (i) ELISA analysis of TNF‐α and IL‐6 in medium supernatant ( n = 3). (j) ELISA analysis of intracellular 2,3‐cGAMP ( n = 3). (k–m) The BMDMs were pre‐transfected with cGAS siRNA or Ns siRNA for 36 h before HR as described in the section of methods. (k) Immunoblotting of cGAS, P‐STING S365, P‐TBK1 S172, TBK1, P‐NF‐κB S536, and GAPDH. (l) ELISA analysis of TNF‐α and IL‐6 in medium supernatant ( n = 3). (m) ELISA analysis of intracellular 2,3‐cGAMP ( n = 3). All these immunoblots have been repeated for 3 times. Data are presented as mean ± SD. * p

Techniques Used: Activation Assay, Enzyme-linked Immunosorbent Assay, Confocal Microscopy, Staining, Fluorescence, Positive Control, Quantitative RT-PCR, Imaging, Transfection, Western Blot

2) Product Images from "Long Non-coding RNA MEG3 Promotes Renal Tubular Epithelial Cell Pyroptosis by Regulating the miR-18a-3p/GSDMD Pathway in Lipopolysaccharide-Induced Acute Kidney Injury"

Article Title: Long Non-coding RNA MEG3 Promotes Renal Tubular Epithelial Cell Pyroptosis by Regulating the miR-18a-3p/GSDMD Pathway in Lipopolysaccharide-Induced Acute Kidney Injury

Journal: Frontiers in Physiology

doi: 10.3389/fphys.2021.663216

Knockdown of MEG3 in TECs inhibited LPS-induced pyroptosis. (A) MEG3 mRNA levels transfected with siRNA. (B) siRNA-transfected TECs stained with EtBr and EthD2. (C) LDH% in supernatants obtained from siRNA-transfected TECs. (D–G) Release of IL-1β and IL-18 in supernatants obtained from siRNA-transfected TECs. (F–H) Protein expression of caspase-1, cleaved caspase-1, GSDMD, and GSDMD-N in siRNA-transfected TECs. ** P
Figure Legend Snippet: Knockdown of MEG3 in TECs inhibited LPS-induced pyroptosis. (A) MEG3 mRNA levels transfected with siRNA. (B) siRNA-transfected TECs stained with EtBr and EthD2. (C) LDH% in supernatants obtained from siRNA-transfected TECs. (D–G) Release of IL-1β and IL-18 in supernatants obtained from siRNA-transfected TECs. (F–H) Protein expression of caspase-1, cleaved caspase-1, GSDMD, and GSDMD-N in siRNA-transfected TECs. ** P

Techniques Used: Transfection, Staining, Expressing

3) Product Images from "Mitochondrial DNA Manipulations Affect Tau Oligomerization"

Article Title: Mitochondrial DNA Manipulations Affect Tau Oligomerization

Journal: Journal of Alzheimer's disease : JAD

doi: 10.3233/JAD-200286

Extent of acute and chronic mtDNA depletion and its effects on respiratory chain protein and mRNA levels. (A) 1, 3, and 7-day EtBr treatment reduced SH-SY5Y mtDNA levels by approximately 50%, 75%, and 95%. ρ0 cells displayed > 99.9% relative mtDNA depletion. (B) Representative western blots labeled with antibodies recognizing the mtDNA-encoded mtCO2, nuclear-encoded COX4I1, and nuclear-encoded NDUFB8 proteins. Actin is included as a loading control. (C) COX4I1 and NDUFB8 mRNA levels were unaffected by the acute EtBr treatment, although the NDUFB8 mRNA level was slightly reduced in ρ0 cells. (D) By MTT assay the acutely mtDNA-depleted cells appeared viable throughout. (E) The acutely mtDNA-depleted cells grossly showed at most subtle morphologic changes. (F) The ρ0 cell cycle rate was slightly reduced as compared to the parent SH-SY5Y line. **p
Figure Legend Snippet: Extent of acute and chronic mtDNA depletion and its effects on respiratory chain protein and mRNA levels. (A) 1, 3, and 7-day EtBr treatment reduced SH-SY5Y mtDNA levels by approximately 50%, 75%, and 95%. ρ0 cells displayed > 99.9% relative mtDNA depletion. (B) Representative western blots labeled with antibodies recognizing the mtDNA-encoded mtCO2, nuclear-encoded COX4I1, and nuclear-encoded NDUFB8 proteins. Actin is included as a loading control. (C) COX4I1 and NDUFB8 mRNA levels were unaffected by the acute EtBr treatment, although the NDUFB8 mRNA level was slightly reduced in ρ0 cells. (D) By MTT assay the acutely mtDNA-depleted cells appeared viable throughout. (E) The acutely mtDNA-depleted cells grossly showed at most subtle morphologic changes. (F) The ρ0 cell cycle rate was slightly reduced as compared to the parent SH-SY5Y line. **p

Techniques Used: Western Blot, Labeling, MTT Assay

Effect of mtDNA depletion on mitochondrial enzyme Vmax activities and energy metabolism fluxes. (A) A progressive decline in complex I and COX activities accompanied the progressive EtBr-induced mtDNA depletion, although after 3 and 7 days of EtBr COX activity fell to a greater extent than complex I activity. CS activity did not decline during this period. (B) SH-SY5Y oxygen consumption rates also tracked mtDNA depletion, and as was the case with the ρ0 cells the mitochondrial OCR was essentially undetectable by EtBr exposure day 7. The EtBr-treated cell ECAR fell initially but recovered, and the EtBr exposure day 7 ECAR exceeded the ρ0 cell ECAR. n=16 per group. (C) Metabolic flux rates for the different groups; the EtBr day 7 cells and ρ0 cells both showed no or essentially no mitochondrial oxygen consumption, but the ρ0 cells uniquely lowered their glycolysis flux. *p
Figure Legend Snippet: Effect of mtDNA depletion on mitochondrial enzyme Vmax activities and energy metabolism fluxes. (A) A progressive decline in complex I and COX activities accompanied the progressive EtBr-induced mtDNA depletion, although after 3 and 7 days of EtBr COX activity fell to a greater extent than complex I activity. CS activity did not decline during this period. (B) SH-SY5Y oxygen consumption rates also tracked mtDNA depletion, and as was the case with the ρ0 cells the mitochondrial OCR was essentially undetectable by EtBr exposure day 7. The EtBr-treated cell ECAR fell initially but recovered, and the EtBr exposure day 7 ECAR exceeded the ρ0 cell ECAR. n=16 per group. (C) Metabolic flux rates for the different groups; the EtBr day 7 cells and ρ0 cells both showed no or essentially no mitochondrial oxygen consumption, but the ρ0 cells uniquely lowered their glycolysis flux. *p

Techniques Used: Activity Assay

Acute and chronic mtDNA depletion alters TOC1 staining. (A) Representative dot blots of EtBr-treated SH-SY5Y lysates labeled with the TOC1 antibody, the Tau12 antibody, or Amido Black. Each day’s EtBR-treated samples were compared to control samples simultaneously prepared on that specific day. Densitometry analysis reveals no change in TOC1 staining after the 1-day EtBr treatment but following 3 and 7 days TOC1 staining increases whether normalized to total protein or total tau. (B) Representative dot blots of SH-SY5Y ρ+ and ρ0 lysates labeled with the TOC1 antibody, the Tau12 antibody, or Amido Black. Densitometry analysis reveals a significant increase in TOC1 staining in ρ0 cells relative to ρ+ cells when normalized to total protein, but not to total tau. **p
Figure Legend Snippet: Acute and chronic mtDNA depletion alters TOC1 staining. (A) Representative dot blots of EtBr-treated SH-SY5Y lysates labeled with the TOC1 antibody, the Tau12 antibody, or Amido Black. Each day’s EtBR-treated samples were compared to control samples simultaneously prepared on that specific day. Densitometry analysis reveals no change in TOC1 staining after the 1-day EtBr treatment but following 3 and 7 days TOC1 staining increases whether normalized to total protein or total tau. (B) Representative dot blots of SH-SY5Y ρ+ and ρ0 lysates labeled with the TOC1 antibody, the Tau12 antibody, or Amido Black. Densitometry analysis reveals a significant increase in TOC1 staining in ρ0 cells relative to ρ+ cells when normalized to total protein, but not to total tau. **p

Techniques Used: Staining, Labeling

Acute mtDNA depletion alters TOC1 staining in differentiated SH-SY5Y cells. (A) Tyrosine hydroxylase protein, a marker of differentiation, increased in staurosporine-treated SH-SY5Y cells. (B) The amount of mtDNA progressively declined in EtBr-treated differentiated cells. (C) Dot blots of differentiated SH-SY5Y cell lysates labeled with the TOC1 antibody, the Tau12 antibody, or Amido Black. Each day’s EtBR-treated samples were compared to control samples simultaneously prepared on that specific day. (D) Densitometry analysis reveals after 1 and 3 days of EtBr, TOC1 staining increases when normalized to total protein. (E) Densitometry analysis reveals after 1 and 3 days of EtBr, TOC1 staining increases when normalized to total tau protein, indicative of a monomer to oligomer shift. n=6 per group. *p
Figure Legend Snippet: Acute mtDNA depletion alters TOC1 staining in differentiated SH-SY5Y cells. (A) Tyrosine hydroxylase protein, a marker of differentiation, increased in staurosporine-treated SH-SY5Y cells. (B) The amount of mtDNA progressively declined in EtBr-treated differentiated cells. (C) Dot blots of differentiated SH-SY5Y cell lysates labeled with the TOC1 antibody, the Tau12 antibody, or Amido Black. Each day’s EtBR-treated samples were compared to control samples simultaneously prepared on that specific day. (D) Densitometry analysis reveals after 1 and 3 days of EtBr, TOC1 staining increases when normalized to total protein. (E) Densitometry analysis reveals after 1 and 3 days of EtBr, TOC1 staining increases when normalized to total tau protein, indicative of a monomer to oligomer shift. n=6 per group. *p

Techniques Used: Staining, Marker, Labeling

4) Product Images from "Anti-Stem Cell Property of Pterostilbene in Gastrointestinal Cancer Cells"

Article Title: Anti-Stem Cell Property of Pterostilbene in Gastrointestinal Cancer Cells

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms21249347

Effect of pterostilbene (PTE) on cell proliferation and expression of stem cell markers in cancer cell lines. ( A ) Cell proliferation was assessed by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay. Cells were treated with PTE for 48 h. ( B ) Apoptosis was assessed by ethidium bromide (EtBr) staining. Insert, CT26 cells treated with PTE for 48 h were stained with EtBr. Arrow head, apoptosis body. Scale bar, 20 µM. ( C ) mRNA expression of stem cell markers; nucleostemin (NS), CD44, Kip2, and CD133 were examined by RT-PCR. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was amplified for loading standard. Lower panels were semi-quantification of stem cell marker expression examined by RT-PCR. Error bar, standard deviation from three independent examinations. Statistical difference was calculated by ordinary analysis of variance. * Statistical difference from PTE (0 µM). PTE, pterostilbene; IC50, 50% inhibitory concentration.
Figure Legend Snippet: Effect of pterostilbene (PTE) on cell proliferation and expression of stem cell markers in cancer cell lines. ( A ) Cell proliferation was assessed by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay. Cells were treated with PTE for 48 h. ( B ) Apoptosis was assessed by ethidium bromide (EtBr) staining. Insert, CT26 cells treated with PTE for 48 h were stained with EtBr. Arrow head, apoptosis body. Scale bar, 20 µM. ( C ) mRNA expression of stem cell markers; nucleostemin (NS), CD44, Kip2, and CD133 were examined by RT-PCR. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was amplified for loading standard. Lower panels were semi-quantification of stem cell marker expression examined by RT-PCR. Error bar, standard deviation from three independent examinations. Statistical difference was calculated by ordinary analysis of variance. * Statistical difference from PTE (0 µM). PTE, pterostilbene; IC50, 50% inhibitory concentration.

Techniques Used: Expressing, MTS Assay, Staining, Reverse Transcription Polymerase Chain Reaction, Amplification, Marker, Standard Deviation, Concentration Assay

5) Product Images from "Antimicrobial and Efflux Pump Inhibitory Activity of Carvotacetones from Sphaeranthus africanus Against Mycobacteria"

Article Title: Antimicrobial and Efflux Pump Inhibitory Activity of Carvotacetones from Sphaeranthus africanus Against Mycobacteria

Journal: Antibiotics

doi: 10.3390/antibiotics9070390

The increase of EtBr accumulation in M . smegmatis mc 2 155 ( a ) and M. aurum ATCC 23366 ( b ) generated by concentrations MIC/2 of the potential efflux pump inhibitors (EPIs) 1 and 6 and the reference inhibitors, verapamil (VP) and chlorpromazine (CPZ), included as two positive controls. EtBr, the negative control, was tested at 0.5 mg/L. The curves represent means ± standard deviations (SD); n = 3.
Figure Legend Snippet: The increase of EtBr accumulation in M . smegmatis mc 2 155 ( a ) and M. aurum ATCC 23366 ( b ) generated by concentrations MIC/2 of the potential efflux pump inhibitors (EPIs) 1 and 6 and the reference inhibitors, verapamil (VP) and chlorpromazine (CPZ), included as two positive controls. EtBr, the negative control, was tested at 0.5 mg/L. The curves represent means ± standard deviations (SD); n = 3.

Techniques Used: Generated, Negative Control

6) Product Images from "Parent and nano-encapsulated ytterbium(iii) complex toward binding with biological macromolecules, in vitro cytotoxicity, cleavage and antimicrobial activity studies †"

Article Title: Parent and nano-encapsulated ytterbium(iii) complex toward binding with biological macromolecules, in vitro cytotoxicity, cleavage and antimicrobial activity studies †

Journal: RSC Advances

doi: 10.1039/d0ra03895d

(A) Fluorescence quenching curves of EtBr bound to DNA by Yb( iii ) complex ([DNA] = 70 μM, [EtBr] = 3.0 μM, [complex] = 0–145.7 μM). Inset is the plot of F / F 0 vs. ([complex]/[DNA]). (B) Fluorescence emission spectra of the mixture solutions of FS-DNA and rhodamine B in the presence of Yb-complex ([Rhodamine B] = 3.0 μM, [complex] = 0–145.7 μM, [FS-DNA] = 70 μM and λ ex = 520).
Figure Legend Snippet: (A) Fluorescence quenching curves of EtBr bound to DNA by Yb( iii ) complex ([DNA] = 70 μM, [EtBr] = 3.0 μM, [complex] = 0–145.7 μM). Inset is the plot of F / F 0 vs. ([complex]/[DNA]). (B) Fluorescence emission spectra of the mixture solutions of FS-DNA and rhodamine B in the presence of Yb-complex ([Rhodamine B] = 3.0 μM, [complex] = 0–145.7 μM, [FS-DNA] = 70 μM and λ ex = 520).

Techniques Used: Fluorescence

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  • 90
    Millipore ao etbr staining
    <t>Apoptosis</t> markers in cancer cells after treatment with CuO NPs. ( A ) Apoptotic indicators for AMJ-13, MCF-7, and HBL-100 cells exposed to CuONPs at IC 50 concentrations for 24 h and stained for 2 min using <t>AO/EtBr.</t> Unexposed cells have an intact structure. Nevertheless, CuONPs exposure correlated with apoptotic aspects indicated using red stains. Scale bar 10 µm. ( B ) Cells cycle phase. Sub-G 1 phase concerning MCF-7 and AMJ-13 cells’ flow cytometry.
    Ao Etbr Staining, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ao etbr staining/product/Millipore
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ao etbr staining - by Bioz Stars, 2022-11
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    90
    Millipore etbr
    <t>Apoptosis</t> markers in cancer cells after treatment with CuO NPs. ( A ) Apoptotic indicators for AMJ-13, MCF-7, and HBL-100 cells exposed to CuONPs at IC 50 concentrations for 24 h and stained for 2 min using <t>AO/EtBr.</t> Unexposed cells have an intact structure. Nevertheless, CuONPs exposure correlated with apoptotic aspects indicated using red stains. Scale bar 10 µm. ( B ) Cells cycle phase. Sub-G 1 phase concerning MCF-7 and AMJ-13 cells’ flow cytometry.
    Etbr, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/etbr/product/Millipore
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    etbr - by Bioz Stars, 2022-11
    90/100 stars
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    88
    Millipore ethidium bromide etbr
    Effects of m AMSA and its acridine derivatives on topoisomerase IIα activity. Effects of derivatives and positive control on the inhibition of human topoisomerase IIα. Native supercoiled pUC19 was incubated for 30 min at 37 °C with 2 units of human Topo IIα in the absence (lane 2) or presence of ligands at a concentration 100 μM. Inhibition of Topo IIα-induced DNA relaxation by m AMSA (lane 3) was used as a positive control. DNA samples were run on an agarose gel followed by <t>ethidium</t> <t>bromide</t> staining.
    Ethidium Bromide Etbr, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Apoptosis markers in cancer cells after treatment with CuO NPs. ( A ) Apoptotic indicators for AMJ-13, MCF-7, and HBL-100 cells exposed to CuONPs at IC 50 concentrations for 24 h and stained for 2 min using AO/EtBr. Unexposed cells have an intact structure. Nevertheless, CuONPs exposure correlated with apoptotic aspects indicated using red stains. Scale bar 10 µm. ( B ) Cells cycle phase. Sub-G 1 phase concerning MCF-7 and AMJ-13 cells’ flow cytometry.

    Journal: Scientific Reports

    Article Title: Biosynthesis of copper oxide nanoparticles mediated Annona muricata as cytotoxic and apoptosis inducer factor in breast cancer cell lines

    doi: 10.1038/s41598-022-20360-y

    Figure Lengend Snippet: Apoptosis markers in cancer cells after treatment with CuO NPs. ( A ) Apoptotic indicators for AMJ-13, MCF-7, and HBL-100 cells exposed to CuONPs at IC 50 concentrations for 24 h and stained for 2 min using AO/EtBr. Unexposed cells have an intact structure. Nevertheless, CuONPs exposure correlated with apoptotic aspects indicated using red stains. Scale bar 10 µm. ( B ) Cells cycle phase. Sub-G 1 phase concerning MCF-7 and AMJ-13 cells’ flow cytometry.

    Article Snippet: Cells treated with copper nanoparticles were assessed for apoptosis induction using AO/EtBr staining (Sigma-Aldrich, USA).

    Techniques: Staining, Flow Cytometry

    Effects of m AMSA and its acridine derivatives on topoisomerase IIα activity. Effects of derivatives and positive control on the inhibition of human topoisomerase IIα. Native supercoiled pUC19 was incubated for 30 min at 37 °C with 2 units of human Topo IIα in the absence (lane 2) or presence of ligands at a concentration 100 μM. Inhibition of Topo IIα-induced DNA relaxation by m AMSA (lane 3) was used as a positive control. DNA samples were run on an agarose gel followed by ethidium bromide staining.

    Journal: Pharmaceuticals

    Article Title: Synthesis and Evaluation of Antiproliferative Activity, Topoisomerase IIα Inhibition, DNA Binding and Non-Clinical Toxicity of New Acridine–Thiosemicarbazone Derivatives

    doi: 10.3390/ph15091098

    Figure Lengend Snippet: Effects of m AMSA and its acridine derivatives on topoisomerase IIα activity. Effects of derivatives and positive control on the inhibition of human topoisomerase IIα. Native supercoiled pUC19 was incubated for 30 min at 37 °C with 2 units of human Topo IIα in the absence (lane 2) or presence of ligands at a concentration 100 μM. Inhibition of Topo IIα-induced DNA relaxation by m AMSA (lane 3) was used as a positive control. DNA samples were run on an agarose gel followed by ethidium bromide staining.

    Article Snippet: Calf thymus DNA (CT DNA), pUC19 DNA plasmid, recombinant human topoisomerase IIα (p170), amsacrine and ethidium bromide (EtBr) used in the interaction analyses were purchased from Sigma-Aldrich.

    Techniques: Activity Assay, Positive Control, Inhibition, Incubation, Concentration Assay, Agarose Gel Electrophoresis, Staining