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monarch gdna binding buffer  (New England Biolabs)


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    New England Biolabs monarch gdna binding buffer
    Monarch Gdna Binding Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/binding+buffer/Monarch+gDNA+Binding+Buffer/10__1016_slash_j__fm__2026__105273-115-4-8
    Average 94 stars, based on 6 article reviews
    monarch gdna binding buffer - by Bioz Stars, 2026-10
    94/100 stars

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    Related Articles

    Binding Assay:

    Article Title: The aging-induced long non-coding RNA MIRIAL controls endothelial cell and mitochondrial function
    Article Snippet: Briefly, HUVECs cultured on 150mm-dishes (Greiner) until confluency, crosslinked with 50 mJ/cm 2 UV light, lyzed in lysis buffer (50 mM Tris/HCl pH 8.0, 150 mM NaCl, 0.5% NP40, 1 mM EDTA, protease inhibitors) and incubated overnight with washed antibody-coated magnetic protein G beads (Dynabeads, invitrogen) in lysis buffer without NP40. .. Beads were then washed with binding buffer (50 mM Tris/HCl pH 8.0, 150 mM NaCl, 0.05% NP40, 1mM EDTA), eluted by proteinase K digest (New England Biolabs) and RNA was recovered using phenol/chloroform/isoamylalcohol (1:1:1) (Roth). .. 5% of the input and the elution fractions were held back, boiled in 4x Laemmli Sample Buffer (Bio-Rad) containing β-mercaptoethanol (AppliChem) and used for subsequent SDS-PAGE and immunoblotting.

    Article Title: Distinct structural and functional heterochromatin partitioning of lamin B1 and B2 revealed using genome-wide Nicking Enzyme Epitope targeted DNA sequencing
    Article Snippet: .. After 3 washes with 1X T-PBS for 5 min at RT, beads were resuspended in 1 ml binding buffer (as described above) including 1 μl of fusion enzyme Nt.CviPII-pGL (NEB) for 1 hr at RT. ..

    Article Title: Meiotic cohesin RAD21L shapes 3D genome structure and transcription in the male germline
    Article Snippet: Beads were then washed twice with 1× binding buffer, resuspended with the dATP attachment mix (1× NEBuffer 2, 0.5 mM dATP, and 2.5 U of NEB Klenow exo minus), and incubated at 37°C for 30 min. .. Subsequently, beads were washed twice with 1× binding buffer, resuspended in 1× NEB Quick ligation reaction buffer together with the NEBNext adapter and Illumina adapters, and incubated 15 min at RT to ligate adapters to chromatin ends. .. Following, beads were washed twice with 1× binding buffer, resuspended with tris buffer, and mixed with 1 μM primer P5, 1 μM primer P7, and cocktail master mix (NEB).

    Article Title: TERRA-LSD1 phase separation promotes R-loop formation for telomere maintenance in ALT cancer cells
    Article Snippet: The supernatant lysates were purified using Ni-NTA Sepharose beads (GE Healthcare). .. After extensive washing with binding buffer, factor Xa protease (New England Biolabs) was incubated with recombinant protein bound to sepharose beads with a concentration of 50 μg ml −1 beads for 4 h at room temperature. ..

    Article Title: Simplified MS2 phage-like particle production including a novel maturation protein internal fusion affinity tag
    Article Snippet: .. The cell pellet was resuspended in 1 mL of Lysis and Binding Buffer (NEB, S1427S) for IMAC or, alternatively, in 1 mL of Buffer W (IBA Lifesciences GmbH, 2-1003-100) for Strep-tag®II (ST) purifications, and lysed by ultrasonic disruption with a Fisherbrand Model 120 Sonic Dismembrator (Thermo Fisher Scientific, FB120A110), while on ice, for 2 total minutes at 40% amplitude with 2 s on and 2 s off, pulse. ..

    Article Title: Road to Extinction? Past and Present Population Structure and Genomic Diversity in the Koala
    Article Snippet: .. The digest was centrifuged, and the supernatant was collected and mixed with 8 × of a binding buffer as detailed in then centrifuged through Monarch DNA Cleanup Columns (5 μg) (New England Biolabs Inc. Beverly, MA, USA). .. DNA bound to the columns was washed with 800 μL buffer PE (Qiagen, Hilden, Germany), then eluted in 25 μL buffer EB (Qiagen).

    Article Title: Simplified MS2 phage-like particle production including a novel maturation protein internal fusion affinity tag
    Article Snippet: .. The cell pellet was resuspended in 1 mL of Lysis and Binding Buffer (NEB, S1427S) for IMAC or, alternatively, in 1 mL of Buffer W (IBA Lifesciences GmbH, 2–1003-100) for Strep-tag®II (ST) purifications, and lysed by ultrasonic disruption with a Fisherbrand Model 120 Sonic Dismembrator (Thermo Fisher Scientific, FB120A110), while on ice, for 2 total minutes at 40% amplitude with 2 s on and 2 s off, pulse. ..

    Article Title: Constitutively bound CTCF sites maintain 3D chromatin architecture and long-range epigenetically regulated domains.
    Article Snippet: Adapters were ligated using the NEBNext Ultra DNA Library Prep kit (#E7370L, NEB). .. Adapter-ligated DNA was washed 2× with 200 μl of 1× tween wash buffer (5mM Tris–HCl pH 8.0, 0.5 mM EDTA, 1M NaCl and 0.05% Tween (#p7949, Sigma Aldrich)), 1× with 200 μl binding buffer and 1× with 200 μl 1× NEB2 buffer (#B7002S, NEB) before beads were resuspended in 30 μl H2O. .. PCR enrichment of adapter-ligated DNA was performed on DNA bound to the MyOne Streptavidin C1 beads using NEB kit (# E7370L, NEB).

    Ligation:

    Article Title: Meiotic cohesin RAD21L shapes 3D genome structure and transcription in the male germline
    Article Snippet: Beads were then washed twice with 1× binding buffer, resuspended with the dATP attachment mix (1× NEBuffer 2, 0.5 mM dATP, and 2.5 U of NEB Klenow exo minus), and incubated at 37°C for 30 min. .. Subsequently, beads were washed twice with 1× binding buffer, resuspended in 1× NEB Quick ligation reaction buffer together with the NEBNext adapter and Illumina adapters, and incubated 15 min at RT to ligate adapters to chromatin ends. .. Following, beads were washed twice with 1× binding buffer, resuspended with tris buffer, and mixed with 1 μM primer P5, 1 μM primer P7, and cocktail master mix (NEB).

    Incubation:

    Article Title: Meiotic cohesin RAD21L shapes 3D genome structure and transcription in the male germline
    Article Snippet: Beads were then washed twice with 1× binding buffer, resuspended with the dATP attachment mix (1× NEBuffer 2, 0.5 mM dATP, and 2.5 U of NEB Klenow exo minus), and incubated at 37°C for 30 min. .. Subsequently, beads were washed twice with 1× binding buffer, resuspended in 1× NEB Quick ligation reaction buffer together with the NEBNext adapter and Illumina adapters, and incubated 15 min at RT to ligate adapters to chromatin ends. .. Following, beads were washed twice with 1× binding buffer, resuspended with tris buffer, and mixed with 1 μM primer P5, 1 μM primer P7, and cocktail master mix (NEB).

    Article Title: TERRA-LSD1 phase separation promotes R-loop formation for telomere maintenance in ALT cancer cells
    Article Snippet: The supernatant lysates were purified using Ni-NTA Sepharose beads (GE Healthcare). .. After extensive washing with binding buffer, factor Xa protease (New England Biolabs) was incubated with recombinant protein bound to sepharose beads with a concentration of 50 μg ml −1 beads for 4 h at room temperature. ..

    Recombinant:

    Article Title: TERRA-LSD1 phase separation promotes R-loop formation for telomere maintenance in ALT cancer cells
    Article Snippet: The supernatant lysates were purified using Ni-NTA Sepharose beads (GE Healthcare). .. After extensive washing with binding buffer, factor Xa protease (New England Biolabs) was incubated with recombinant protein bound to sepharose beads with a concentration of 50 μg ml −1 beads for 4 h at room temperature. ..

    Concentration Assay:

    Article Title: TERRA-LSD1 phase separation promotes R-loop formation for telomere maintenance in ALT cancer cells
    Article Snippet: The supernatant lysates were purified using Ni-NTA Sepharose beads (GE Healthcare). .. After extensive washing with binding buffer, factor Xa protease (New England Biolabs) was incubated with recombinant protein bound to sepharose beads with a concentration of 50 μg ml −1 beads for 4 h at room temperature. ..

    Lysis:

    Article Title: Simplified MS2 phage-like particle production including a novel maturation protein internal fusion affinity tag
    Article Snippet: .. The cell pellet was resuspended in 1 mL of Lysis and Binding Buffer (NEB, S1427S) for IMAC or, alternatively, in 1 mL of Buffer W (IBA Lifesciences GmbH, 2-1003-100) for Strep-tag®II (ST) purifications, and lysed by ultrasonic disruption with a Fisherbrand Model 120 Sonic Dismembrator (Thermo Fisher Scientific, FB120A110), while on ice, for 2 total minutes at 40% amplitude with 2 s on and 2 s off, pulse. ..

    Article Title: Simplified MS2 phage-like particle production including a novel maturation protein internal fusion affinity tag
    Article Snippet: .. The cell pellet was resuspended in 1 mL of Lysis and Binding Buffer (NEB, S1427S) for IMAC or, alternatively, in 1 mL of Buffer W (IBA Lifesciences GmbH, 2–1003-100) for Strep-tag®II (ST) purifications, and lysed by ultrasonic disruption with a Fisherbrand Model 120 Sonic Dismembrator (Thermo Fisher Scientific, FB120A110), while on ice, for 2 total minutes at 40% amplitude with 2 s on and 2 s off, pulse. ..

    Disruption:

    Article Title: Simplified MS2 phage-like particle production including a novel maturation protein internal fusion affinity tag
    Article Snippet: .. The cell pellet was resuspended in 1 mL of Lysis and Binding Buffer (NEB, S1427S) for IMAC or, alternatively, in 1 mL of Buffer W (IBA Lifesciences GmbH, 2-1003-100) for Strep-tag®II (ST) purifications, and lysed by ultrasonic disruption with a Fisherbrand Model 120 Sonic Dismembrator (Thermo Fisher Scientific, FB120A110), while on ice, for 2 total minutes at 40% amplitude with 2 s on and 2 s off, pulse. ..

    Article Title: Simplified MS2 phage-like particle production including a novel maturation protein internal fusion affinity tag
    Article Snippet: .. The cell pellet was resuspended in 1 mL of Lysis and Binding Buffer (NEB, S1427S) for IMAC or, alternatively, in 1 mL of Buffer W (IBA Lifesciences GmbH, 2–1003-100) for Strep-tag®II (ST) purifications, and lysed by ultrasonic disruption with a Fisherbrand Model 120 Sonic Dismembrator (Thermo Fisher Scientific, FB120A110), while on ice, for 2 total minutes at 40% amplitude with 2 s on and 2 s off, pulse. ..



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    Image Search Results


    ( A ) Immunoblotting of SIINFEKL peptide–pulsed Neuro2a cell lines cocultured with activated OT-I T cells for 4 hours at ratios of T cells to target cells indicated. ( B ) Schematic representation of fluorescence-activated cell sorting (FACS) based on Caspase-3(DEVD)NucView and annexin V (AnnV) staining, applies to (C) to (E). Created in BioRender; B. Liu (2026), https://BioRender.com/y3nlv0g . ( C ) Rate of outgrowth from cells recovered from FACS sort, calculated on the basis of number of initially plated cells. Data were compiled from five independent experiments. Paired t test. ( D and E ) Quantification of nucleotide fragments with greater than three, four, five, or six single-nucleotide variants (SNVs) from various recovered populations as indicated. ( F ) The design of the ATM–fluorescence resonance energy transfer (FRET) reporter. Created in BioRender; B. Liu (2026), https://BioRender.com/jqdhdx7 . ( G ) Quantification of FRET loss signal from Neuro2a expressing the ATM-FRET reporter shown in (F). Neuro2a cells were SIINFEKL peptide pulsed and then coincubated with OT-I T cells for 6 hours at 10:1 T cell–to–target cell ratio. Data were pooled from three independent experiments. Unpaired t test. ( H ) In vivo growth of Neuro2a cells expressing the ATM-FRET sensor. ATM-FRET loss populations were sorted post BH3m treatment (1 μM ABT737 and 1 μM S63845 ). Cells were sorted for FRET-loss population at 2.5 hours, and 1 × 10 4 cells were injected into NSG mice ( n = 5 animals per group). Kaplan-Meyer survival curves. WT-base and GE KO–base refer to ATM-FRET–expressing cells that were not treated with BH3m. Log-rank (Mantel-Cox) test. * P < 0.05 and **** P < 0.0001. C3, active caspase-3.

    Journal: Science Advances

    Article Title: Tumor gasdermin E acts as a rheostat to limit apoptotic cell survival and mutagenesis in anticancer immune environments

    doi: 10.1126/sciadv.aeb1137

    Figure Lengend Snippet: ( A ) Immunoblotting of SIINFEKL peptide–pulsed Neuro2a cell lines cocultured with activated OT-I T cells for 4 hours at ratios of T cells to target cells indicated. ( B ) Schematic representation of fluorescence-activated cell sorting (FACS) based on Caspase-3(DEVD)NucView and annexin V (AnnV) staining, applies to (C) to (E). Created in BioRender; B. Liu (2026), https://BioRender.com/y3nlv0g . ( C ) Rate of outgrowth from cells recovered from FACS sort, calculated on the basis of number of initially plated cells. Data were compiled from five independent experiments. Paired t test. ( D and E ) Quantification of nucleotide fragments with greater than three, four, five, or six single-nucleotide variants (SNVs) from various recovered populations as indicated. ( F ) The design of the ATM–fluorescence resonance energy transfer (FRET) reporter. Created in BioRender; B. Liu (2026), https://BioRender.com/jqdhdx7 . ( G ) Quantification of FRET loss signal from Neuro2a expressing the ATM-FRET reporter shown in (F). Neuro2a cells were SIINFEKL peptide pulsed and then coincubated with OT-I T cells for 6 hours at 10:1 T cell–to–target cell ratio. Data were pooled from three independent experiments. Unpaired t test. ( H ) In vivo growth of Neuro2a cells expressing the ATM-FRET sensor. ATM-FRET loss populations were sorted post BH3m treatment (1 μM ABT737 and 1 μM S63845 ). Cells were sorted for FRET-loss population at 2.5 hours, and 1 × 10 4 cells were injected into NSG mice ( n = 5 animals per group). Kaplan-Meyer survival curves. WT-base and GE KO–base refer to ATM-FRET–expressing cells that were not treated with BH3m. Log-rank (Mantel-Cox) test. * P < 0.05 and **** P < 0.0001. C3, active caspase-3.

    Article Snippet: Cells were spun down and resuspended in DMEM supplemented with annexin binding buffer (99902, Biotium) and stained with annexin V-647 and/or Caspase-3(DEVD)NucView-488, as indicated in the text, for 10 min at 37°C.

    Techniques: Western Blot, Fluorescence, FACS, Staining, Förster Resonance Energy Transfer, Expressing, In Vivo, Injection

    ( A ) Coculture (1:1) of parental B16F10 cells expressing pBabe-Cerulean (blue) and B16F10 cells expressing GsdmD-C3 and mCherry (red) (see movie S1). ( B ) Times of individual cell shrinkage and cell membrane lysis, as in (A). Unpaired t test. ( C ) Apoptotic phase (shrinkage-lysis interval) from (B). One-way ANOVA. ( D ) Quantification of annexin V + SIINFEKL peptide–pulsed B16F10 [as in (B) and (C)] that remains fluorescent when coincubated with activated OT-I T cells at 4 hours. Two-way ANOVA. ( E ) Quantification of SIINFEKL peptide–pulsed B16F10 cell populations with ectopic caspase-3-activatable GSDMD (GD-C3) by fluorescence signal at 24 hours of coincubation with activated OT-I T cells. ( F ) As in (E) with Gsdme. [(E) and (F)] Paired t test. ( G ) Number of GCamp6s spikes observed from SIINFEKL peptide–pulsed Neuro2a cells, coincubated with activated OT-I T cells, 10:1 T cell–to–target cell ratio. Images were taken every 2 min for a duration of 3 hours immediately following the additional of OT-I T cells, and numbers of observable GCamp6s spikes were counted and plotted (see movies S2 to S4 and fig. S5C). Unpaired t test. ( H ) Design for (I) and (J). Created in BioRender; B. Liu (2026), https://BioRender.com/j67ipmw . ( I ) Bioluminescence signal intensity of Neuro2a in C57BL/6 animals ( n = 9 animals per group). Two-way ANOVA. ( J ) Kaplan-Meyer survival curves from C57BL/6 mice injected with Neuro2a cells ( n = 9 animals per group). ( K ) Design for (L). Created in BioRender; B. Liu (2026), https://BioRender.com/j67ipmw . ( L ) Bioluminescence signal intensity of Neuro2a in C57BL/6 animals (control, n = 9 animals; Gsdme −/− , n = 8 animals). ( M ) Kaplan-Meyer survival curves from C57BL/6 mice rechallenged with Neuro2a cells [ n = 5 animals per group; log-rank (Mantel-Cox) test]. ( N ) As in (M) with B16F10 [ n = 12 animals in naïve group, 14 animals in rechallenge group; log-rank (Mantel-Cox) test]. * P < 0.05, ** P < 0.01, and **** P < 0.0001. iv, intravenous; ip, intraperitoneal.

    Journal: Science Advances

    Article Title: Tumor gasdermin E acts as a rheostat to limit apoptotic cell survival and mutagenesis in anticancer immune environments

    doi: 10.1126/sciadv.aeb1137

    Figure Lengend Snippet: ( A ) Coculture (1:1) of parental B16F10 cells expressing pBabe-Cerulean (blue) and B16F10 cells expressing GsdmD-C3 and mCherry (red) (see movie S1). ( B ) Times of individual cell shrinkage and cell membrane lysis, as in (A). Unpaired t test. ( C ) Apoptotic phase (shrinkage-lysis interval) from (B). One-way ANOVA. ( D ) Quantification of annexin V + SIINFEKL peptide–pulsed B16F10 [as in (B) and (C)] that remains fluorescent when coincubated with activated OT-I T cells at 4 hours. Two-way ANOVA. ( E ) Quantification of SIINFEKL peptide–pulsed B16F10 cell populations with ectopic caspase-3-activatable GSDMD (GD-C3) by fluorescence signal at 24 hours of coincubation with activated OT-I T cells. ( F ) As in (E) with Gsdme. [(E) and (F)] Paired t test. ( G ) Number of GCamp6s spikes observed from SIINFEKL peptide–pulsed Neuro2a cells, coincubated with activated OT-I T cells, 10:1 T cell–to–target cell ratio. Images were taken every 2 min for a duration of 3 hours immediately following the additional of OT-I T cells, and numbers of observable GCamp6s spikes were counted and plotted (see movies S2 to S4 and fig. S5C). Unpaired t test. ( H ) Design for (I) and (J). Created in BioRender; B. Liu (2026), https://BioRender.com/j67ipmw . ( I ) Bioluminescence signal intensity of Neuro2a in C57BL/6 animals ( n = 9 animals per group). Two-way ANOVA. ( J ) Kaplan-Meyer survival curves from C57BL/6 mice injected with Neuro2a cells ( n = 9 animals per group). ( K ) Design for (L). Created in BioRender; B. Liu (2026), https://BioRender.com/j67ipmw . ( L ) Bioluminescence signal intensity of Neuro2a in C57BL/6 animals (control, n = 9 animals; Gsdme −/− , n = 8 animals). ( M ) Kaplan-Meyer survival curves from C57BL/6 mice rechallenged with Neuro2a cells [ n = 5 animals per group; log-rank (Mantel-Cox) test]. ( N ) As in (M) with B16F10 [ n = 12 animals in naïve group, 14 animals in rechallenge group; log-rank (Mantel-Cox) test]. * P < 0.05, ** P < 0.01, and **** P < 0.0001. iv, intravenous; ip, intraperitoneal.

    Article Snippet: Cells were spun down and resuspended in DMEM supplemented with annexin binding buffer (99902, Biotium) and stained with annexin V-647 and/or Caspase-3(DEVD)NucView-488, as indicated in the text, for 10 min at 37°C.

    Techniques: Expressing, Membrane, Lysis, Fluorescence, Injection, Control

    CRISPR/Cas9 induces DNA damage and apoptosis in PGCs. (A) Flow cytometry analysis 24 h after electroporation, quantifying the proportion of Annexin V + /PI + cells. The horizontal axis indicates PI and the vertical axis Annexin V. The upper-left quadrant (Annexin V + /PI + ) represents late apoptotic cells, and the lower-right quadrant (Annexin V + only) represents early apoptotic cells. Upper panels: results after electroporation with Cas9 + various gRNAs; lower panels: results with dCas9 + various gRNAs. (B) Bar graph of Annexin V + /PI + percentages across groups. Cas9 editing induced a highly significant increase in late apoptosis. (C) γ-H 2 AX foci (green) detected by immunofluorescence 24 h after electroporation. Foci appear as discrete nuclear puncta; nuclei are counterstained with DAPI (blue). Scale bar = 10 µm. (D) Quantification of γ-H 2 AX foci per cell. Cas9 targeting resulted in a significant increase in γ-H 2 AX foci per cell, whereas dCas9 with sgRNA did not. Statistical significance determined by one-way ANOVA (p-values are indicated in the figure).

    Journal: Poultry Science

    Article Title: High genotoxicity of CRISPR/Cas9 versus limited efficacy of CRISPRi in chicken primordial germ cells

    doi: 10.1016/j.psj.2026.106722

    Figure Lengend Snippet: CRISPR/Cas9 induces DNA damage and apoptosis in PGCs. (A) Flow cytometry analysis 24 h after electroporation, quantifying the proportion of Annexin V + /PI + cells. The horizontal axis indicates PI and the vertical axis Annexin V. The upper-left quadrant (Annexin V + /PI + ) represents late apoptotic cells, and the lower-right quadrant (Annexin V + only) represents early apoptotic cells. Upper panels: results after electroporation with Cas9 + various gRNAs; lower panels: results with dCas9 + various gRNAs. (B) Bar graph of Annexin V + /PI + percentages across groups. Cas9 editing induced a highly significant increase in late apoptosis. (C) γ-H 2 AX foci (green) detected by immunofluorescence 24 h after electroporation. Foci appear as discrete nuclear puncta; nuclei are counterstained with DAPI (blue). Scale bar = 10 µm. (D) Quantification of γ-H 2 AX foci per cell. Cas9 targeting resulted in a significant increase in γ-H 2 AX foci per cell, whereas dCas9 with sgRNA did not. Statistical significance determined by one-way ANOVA (p-values are indicated in the figure).

    Article Snippet: After another centrifugation, the supernatant was removed and cells were resuspended in 500 μL of 1× Annexin V Binding Buffer (from Annexin V-FITC/PI Apoptosis Kit, Elabscience E-CK-A211).

    Techniques: CRISPR, Flow Cytometry, Electroporation, Immunofluorescence

    PGCs are more sensitive to DNA double-strand breaks than somatic cells. (A) Cell viability curves after 24 h treatment with increasing concentrations of etoposide (ETP; x-axis shows log 10 [ETP]). PGC viability dropped sharply even at low ETP doses, whereas CEF cells were more tolerant (viability data are mean ± SD of triplicates). (B) Apoptosis detection in PGCs treated with low-dose ETP. The proportion of Annexin V + /PI + cells (late apoptosis) was significantly elevated even at 0.03 µM ETP. (C) Western blot analysis of γ-H 2 AX protein in PGCs treated with varying ETP concentrations (left panel; β-actin as loading control). The bar graph (right) shows the ratio of γ-H 2 AX to β-actin band intensity, with a marked increase at 3 µM ETP (p-values are indicated in the figure). (D) Quantification of γ-H 2 AX foci per cell in THP-1 cells, male PGCs, and female PGCs after exposure to X-ray doses of 0, 2, 4, 6, and 8 Gy. After 48 h recovery, γ-H 2 AX foci increased significantly with higher radiation in THP-1, male PGCs, and female PGCs. Female PGC data points are red squares; male PGCs are blue squares; THP-1 are black circles. (E) Cell-cycle distribution of male vs. female PGCs after DNA damage. PGCs were irradiated (2, 4, 6 Gy), cultured 48 h, and analyzed by flow cytometry for cell-cycle phase (propidium iodide staining). Stacked bars show the percentage of cells in G 0 G 1 , S, and G 2 /M phases in untreated vs. irradiated cells. After damage, female PGCs predominantly arrested in G 2 /M (increased G 2 fraction), whereas male PGCs accumulated in S phase. Statistical significance by one-way ANOVA (p-values are indicated in the figure).

    Journal: Poultry Science

    Article Title: High genotoxicity of CRISPR/Cas9 versus limited efficacy of CRISPRi in chicken primordial germ cells

    doi: 10.1016/j.psj.2026.106722

    Figure Lengend Snippet: PGCs are more sensitive to DNA double-strand breaks than somatic cells. (A) Cell viability curves after 24 h treatment with increasing concentrations of etoposide (ETP; x-axis shows log 10 [ETP]). PGC viability dropped sharply even at low ETP doses, whereas CEF cells were more tolerant (viability data are mean ± SD of triplicates). (B) Apoptosis detection in PGCs treated with low-dose ETP. The proportion of Annexin V + /PI + cells (late apoptosis) was significantly elevated even at 0.03 µM ETP. (C) Western blot analysis of γ-H 2 AX protein in PGCs treated with varying ETP concentrations (left panel; β-actin as loading control). The bar graph (right) shows the ratio of γ-H 2 AX to β-actin band intensity, with a marked increase at 3 µM ETP (p-values are indicated in the figure). (D) Quantification of γ-H 2 AX foci per cell in THP-1 cells, male PGCs, and female PGCs after exposure to X-ray doses of 0, 2, 4, 6, and 8 Gy. After 48 h recovery, γ-H 2 AX foci increased significantly with higher radiation in THP-1, male PGCs, and female PGCs. Female PGC data points are red squares; male PGCs are blue squares; THP-1 are black circles. (E) Cell-cycle distribution of male vs. female PGCs after DNA damage. PGCs were irradiated (2, 4, 6 Gy), cultured 48 h, and analyzed by flow cytometry for cell-cycle phase (propidium iodide staining). Stacked bars show the percentage of cells in G 0 G 1 , S, and G 2 /M phases in untreated vs. irradiated cells. After damage, female PGCs predominantly arrested in G 2 /M (increased G 2 fraction), whereas male PGCs accumulated in S phase. Statistical significance by one-way ANOVA (p-values are indicated in the figure).

    Article Snippet: After another centrifugation, the supernatant was removed and cells were resuspended in 500 μL of 1× Annexin V Binding Buffer (from Annexin V-FITC/PI Apoptosis Kit, Elabscience E-CK-A211).

    Techniques: Western Blot, Control, Irradiation, Cell Culture, Flow Cytometry, Staining