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mouse anti-trf2 antibody 4a794  (Millipore)


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    Structured Review

    Millipore mouse anti-trf2 antibody 4a794
    Mouse Anti Trf2 Antibody 4a794, 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/product/mouse+anti-trf2+antibody/anti+trf2/pm36426578-350-7-10
    Average 90 stars, based on 1 article reviews
    mouse anti-trf2 antibody 4a794 - by Bioz Stars, 2026-09
    90/100 stars

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    Related Articles

    Incubation:

    Article Title: MiR ‐182‐3p targets TRF2 and impairs tumor growth of triple‐negative breast cancer
    Article Snippet: For immune labeling, cells were incubated with mouse anti-TRF2 (1:2,000, Millipore, 4A794) antibody in 3% BSA, 0.1% Tween-20 (1× PBS) for 1 h at RT.

    Article Title: The structurally conserved TELR region on shelterin protein TPP1 is essential for telomerase processivity but not recruitment
    Article Snippet: Immunofluorescence staining and FISH were carried out as described previously ( ) with the following antibodies: rabbit anti-GFP antibody (Novus; NB600-308), mouse anti-TRF1 antibody (GeneTex; GTX70304). mouse anti-TRF2 antibody (Millipore; 05-521), rabbit anti-coilin antibody (GeneTex; GTX112570).

    Article Title: Recruitment of TRF2 to laser-induced DNA damage sites.
    Article Snippet: Several lines of evidence suggest that the telomere-associated protein TRF2 plays critical roles in the DNA damage response.. TRF2 is rapidly and transiently phosphorylated by an ATM-dependent pathway in response to DNA damage and this DNA damage-induced phosphoryation is essential for the DNA-PKdependent pathway of DNA double-strand break repair (DSB).. However, the type of DNA damage that induces TRF2 localization to the damage sites, the requirement for DNA damage-induced phosphorylation of TRF2 for its recruitment, as well as the detailed kinetics of TRF2 accumulation at DNA damage sites have not been fully investigated.

    Article Title: MiR-182-3p targets TRF2 and impairs tumor growth of triple-negative breast cancer.
    Article Snippet: The telomeric repeat-binding factor 2 (TRF2) is a telomere-capping protein that plays a key role in the maintenance of telomere structure and function.. It is highly expressed in different cancer types, and it contributes to cancer progression.. To date, anti-cancer strategies to target TRF2 remain a challenge.

    Labeling:

    Article Title: MiR ‐182‐3p targets TRF2 and impairs tumor growth of triple‐negative breast cancer
    Article Snippet: For immune labeling, cells were incubated with mouse anti-TRF2 (1:2,000, Millipore, 4A794) antibody in 3% BSA, 0.1% Tween-20 (1× PBS) for 1 h at RT.

    Article Title: The structurally conserved TELR region on shelterin protein TPP1 is essential for telomerase processivity but not recruitment
    Article Snippet: Immunofluorescence staining and FISH were carried out as described previously ( ) with the following antibodies: rabbit anti-GFP antibody (Novus; NB600-308), mouse anti-TRF1 antibody (GeneTex; GTX70304). mouse anti-TRF2 antibody (Millipore; 05-521), rabbit anti-coilin antibody (GeneTex; GTX112570).

    Article Title: Recruitment of TRF2 to laser-induced DNA damage sites.
    Article Snippet: Several lines of evidence suggest that the telomere-associated protein TRF2 plays critical roles in the DNA damage response.. TRF2 is rapidly and transiently phosphorylated by an ATM-dependent pathway in response to DNA damage and this DNA damage-induced phosphoryation is essential for the DNA-PKdependent pathway of DNA double-strand break repair (DSB).. However, the type of DNA damage that induces TRF2 localization to the damage sites, the requirement for DNA damage-induced phosphorylation of TRF2 for its recruitment, as well as the detailed kinetics of TRF2 accumulation at DNA damage sites have not been fully investigated.

    Article Title: MiR-182-3p targets TRF2 and impairs tumor growth of triple-negative breast cancer.
    Article Snippet: The telomeric repeat-binding factor 2 (TRF2) is a telomere-capping protein that plays a key role in the maintenance of telomere structure and function.. It is highly expressed in different cancer types, and it contributes to cancer progression.. To date, anti-cancer strategies to target TRF2 remain a challenge.

    Immunofluorescence:

    Article Title: MiR ‐182‐3p targets TRF2 and impairs tumor growth of triple‐negative breast cancer
    Article Snippet: For immune labeling, cells were incubated with mouse anti-TRF2 (1:2,000, Millipore, 4A794) antibody in 3% BSA, 0.1% Tween-20 (1× PBS) for 1 h at RT.

    Article Title: The structurally conserved TELR region on shelterin protein TPP1 is essential for telomerase processivity but not recruitment
    Article Snippet: Immunofluorescence staining and FISH were carried out as described previously ( ) with the following antibodies: rabbit anti-GFP antibody (Novus; NB600-308), mouse anti-TRF1 antibody (GeneTex; GTX70304). mouse anti-TRF2 antibody (Millipore; 05-521), rabbit anti-coilin antibody (GeneTex; GTX112570).

    Article Title: Recruitment of TRF2 to laser-induced DNA damage sites.
    Article Snippet: Several lines of evidence suggest that the telomere-associated protein TRF2 plays critical roles in the DNA damage response.. TRF2 is rapidly and transiently phosphorylated by an ATM-dependent pathway in response to DNA damage and this DNA damage-induced phosphoryation is essential for the DNA-PKdependent pathway of DNA double-strand break repair (DSB).. However, the type of DNA damage that induces TRF2 localization to the damage sites, the requirement for DNA damage-induced phosphorylation of TRF2 for its recruitment, as well as the detailed kinetics of TRF2 accumulation at DNA damage sites have not been fully investigated.

    Article Title: MiR-182-3p targets TRF2 and impairs tumor growth of triple-negative breast cancer.
    Article Snippet: The telomeric repeat-binding factor 2 (TRF2) is a telomere-capping protein that plays a key role in the maintenance of telomere structure and function.. It is highly expressed in different cancer types, and it contributes to cancer progression.. To date, anti-cancer strategies to target TRF2 remain a challenge.

    Staining:

    Article Title: MiR ‐182‐3p targets TRF2 and impairs tumor growth of triple‐negative breast cancer
    Article Snippet: For immune labeling, cells were incubated with mouse anti-TRF2 (1:2,000, Millipore, 4A794) antibody in 3% BSA, 0.1% Tween-20 (1× PBS) for 1 h at RT.

    Article Title: The structurally conserved TELR region on shelterin protein TPP1 is essential for telomerase processivity but not recruitment
    Article Snippet: Immunofluorescence staining and FISH were carried out as described previously ( ) with the following antibodies: rabbit anti-GFP antibody (Novus; NB600-308), mouse anti-TRF1 antibody (GeneTex; GTX70304). mouse anti-TRF2 antibody (Millipore; 05-521), rabbit anti-coilin antibody (GeneTex; GTX112570).

    Article Title: Recruitment of TRF2 to laser-induced DNA damage sites.
    Article Snippet: Several lines of evidence suggest that the telomere-associated protein TRF2 plays critical roles in the DNA damage response.. TRF2 is rapidly and transiently phosphorylated by an ATM-dependent pathway in response to DNA damage and this DNA damage-induced phosphoryation is essential for the DNA-PKdependent pathway of DNA double-strand break repair (DSB).. However, the type of DNA damage that induces TRF2 localization to the damage sites, the requirement for DNA damage-induced phosphorylation of TRF2 for its recruitment, as well as the detailed kinetics of TRF2 accumulation at DNA damage sites have not been fully investigated.

    Article Title: MiR-182-3p targets TRF2 and impairs tumor growth of triple-negative breast cancer.
    Article Snippet: The telomeric repeat-binding factor 2 (TRF2) is a telomere-capping protein that plays a key role in the maintenance of telomere structure and function.. It is highly expressed in different cancer types, and it contributes to cancer progression.. To date, anti-cancer strategies to target TRF2 remain a challenge.

    Fluorescence In Situ Hybridization:

    Article Title: MiR ‐182‐3p targets TRF2 and impairs tumor growth of triple‐negative breast cancer
    Article Snippet: For immune labeling, cells were incubated with mouse anti-TRF2 (1:2,000, Millipore, 4A794) antibody in 3% BSA, 0.1% Tween-20 (1× PBS) for 1 h at RT.

    Article Title: The structurally conserved TELR region on shelterin protein TPP1 is essential for telomerase processivity but not recruitment
    Article Snippet: Immunofluorescence staining and FISH were carried out as described previously ( ) with the following antibodies: rabbit anti-GFP antibody (Novus; NB600-308), mouse anti-TRF1 antibody (GeneTex; GTX70304). mouse anti-TRF2 antibody (Millipore; 05-521), rabbit anti-coilin antibody (GeneTex; GTX112570).

    Article Title: Recruitment of TRF2 to laser-induced DNA damage sites.
    Article Snippet: Several lines of evidence suggest that the telomere-associated protein TRF2 plays critical roles in the DNA damage response.. TRF2 is rapidly and transiently phosphorylated by an ATM-dependent pathway in response to DNA damage and this DNA damage-induced phosphoryation is essential for the DNA-PKdependent pathway of DNA double-strand break repair (DSB).. However, the type of DNA damage that induces TRF2 localization to the damage sites, the requirement for DNA damage-induced phosphorylation of TRF2 for its recruitment, as well as the detailed kinetics of TRF2 accumulation at DNA damage sites have not been fully investigated.

    Article Title: MiR-182-3p targets TRF2 and impairs tumor growth of triple-negative breast cancer.
    Article Snippet: The telomeric repeat-binding factor 2 (TRF2) is a telomere-capping protein that plays a key role in the maintenance of telomere structure and function.. It is highly expressed in different cancer types, and it contributes to cancer progression.. To date, anti-cancer strategies to target TRF2 remain a challenge.

    Transfection:

    Article Title: MiR ‐182‐3p targets TRF2 and impairs tumor growth of triple‐negative breast cancer
    Article Snippet: For immune labeling, cells were incubated with mouse anti-TRF2 (1:2,000, Millipore, 4A794) antibody in 3% BSA, 0.1% Tween-20 (1× PBS) for 1 h at RT.

    Article Title: The structurally conserved TELR region on shelterin protein TPP1 is essential for telomerase processivity but not recruitment
    Article Snippet: Immunofluorescence staining and FISH were carried out as described previously ( ) with the following antibodies: rabbit anti-GFP antibody (Novus; NB600-308), mouse anti-TRF1 antibody (GeneTex; GTX70304). mouse anti-TRF2 antibody (Millipore; 05-521), rabbit anti-coilin antibody (GeneTex; GTX112570).

    Article Title: Recruitment of TRF2 to laser-induced DNA damage sites.
    Article Snippet: Several lines of evidence suggest that the telomere-associated protein TRF2 plays critical roles in the DNA damage response.. TRF2 is rapidly and transiently phosphorylated by an ATM-dependent pathway in response to DNA damage and this DNA damage-induced phosphoryation is essential for the DNA-PKdependent pathway of DNA double-strand break repair (DSB).. However, the type of DNA damage that induces TRF2 localization to the damage sites, the requirement for DNA damage-induced phosphorylation of TRF2 for its recruitment, as well as the detailed kinetics of TRF2 accumulation at DNA damage sites have not been fully investigated.

    Article Title: MiR-182-3p targets TRF2 and impairs tumor growth of triple-negative breast cancer.
    Article Snippet: The telomeric repeat-binding factor 2 (TRF2) is a telomere-capping protein that plays a key role in the maintenance of telomere structure and function.. It is highly expressed in different cancer types, and it contributes to cancer progression.. To date, anti-cancer strategies to target TRF2 remain a challenge.

    Expressing:

    Article Title: MiR ‐182‐3p targets TRF2 and impairs tumor growth of triple‐negative breast cancer
    Article Snippet: For immune labeling, cells were incubated with mouse anti-TRF2 (1:2,000, Millipore, 4A794) antibody in 3% BSA, 0.1% Tween-20 (1× PBS) for 1 h at RT.

    Article Title: The structurally conserved TELR region on shelterin protein TPP1 is essential for telomerase processivity but not recruitment
    Article Snippet: Immunofluorescence staining and FISH were carried out as described previously ( ) with the following antibodies: rabbit anti-GFP antibody (Novus; NB600-308), mouse anti-TRF1 antibody (GeneTex; GTX70304). mouse anti-TRF2 antibody (Millipore; 05-521), rabbit anti-coilin antibody (GeneTex; GTX112570).

    Article Title: Recruitment of TRF2 to laser-induced DNA damage sites.
    Article Snippet: Several lines of evidence suggest that the telomere-associated protein TRF2 plays critical roles in the DNA damage response.. TRF2 is rapidly and transiently phosphorylated by an ATM-dependent pathway in response to DNA damage and this DNA damage-induced phosphoryation is essential for the DNA-PKdependent pathway of DNA double-strand break repair (DSB).. However, the type of DNA damage that induces TRF2 localization to the damage sites, the requirement for DNA damage-induced phosphorylation of TRF2 for its recruitment, as well as the detailed kinetics of TRF2 accumulation at DNA damage sites have not been fully investigated.

    Article Title: MiR-182-3p targets TRF2 and impairs tumor growth of triple-negative breast cancer.
    Article Snippet: The telomeric repeat-binding factor 2 (TRF2) is a telomere-capping protein that plays a key role in the maintenance of telomere structure and function.. It is highly expressed in different cancer types, and it contributes to cancer progression.. To date, anti-cancer strategies to target TRF2 remain a challenge.

    Construct:

    Article Title: MiR ‐182‐3p targets TRF2 and impairs tumor growth of triple‐negative breast cancer
    Article Snippet: For immune labeling, cells were incubated with mouse anti-TRF2 (1:2,000, Millipore, 4A794) antibody in 3% BSA, 0.1% Tween-20 (1× PBS) for 1 h at RT.

    Article Title: The structurally conserved TELR region on shelterin protein TPP1 is essential for telomerase processivity but not recruitment
    Article Snippet: Immunofluorescence staining and FISH were carried out as described previously ( ) with the following antibodies: rabbit anti-GFP antibody (Novus; NB600-308), mouse anti-TRF1 antibody (GeneTex; GTX70304). mouse anti-TRF2 antibody (Millipore; 05-521), rabbit anti-coilin antibody (GeneTex; GTX112570).

    Article Title: Recruitment of TRF2 to laser-induced DNA damage sites.
    Article Snippet: Several lines of evidence suggest that the telomere-associated protein TRF2 plays critical roles in the DNA damage response.. TRF2 is rapidly and transiently phosphorylated by an ATM-dependent pathway in response to DNA damage and this DNA damage-induced phosphoryation is essential for the DNA-PKdependent pathway of DNA double-strand break repair (DSB).. However, the type of DNA damage that induces TRF2 localization to the damage sites, the requirement for DNA damage-induced phosphorylation of TRF2 for its recruitment, as well as the detailed kinetics of TRF2 accumulation at DNA damage sites have not been fully investigated.

    Article Title: MiR-182-3p targets TRF2 and impairs tumor growth of triple-negative breast cancer.
    Article Snippet: The telomeric repeat-binding factor 2 (TRF2) is a telomere-capping protein that plays a key role in the maintenance of telomere structure and function.. It is highly expressed in different cancer types, and it contributes to cancer progression.. To date, anti-cancer strategies to target TRF2 remain a challenge.

    Immunostaining:

    Article Title: MiR ‐182‐3p targets TRF2 and impairs tumor growth of triple‐negative breast cancer
    Article Snippet: For immune labeling, cells were incubated with mouse anti-TRF2 (1:2,000, Millipore, 4A794) antibody in 3% BSA, 0.1% Tween-20 (1× PBS) for 1 h at RT.

    Article Title: The structurally conserved TELR region on shelterin protein TPP1 is essential for telomerase processivity but not recruitment
    Article Snippet: Immunofluorescence staining and FISH were carried out as described previously ( ) with the following antibodies: rabbit anti-GFP antibody (Novus; NB600-308), mouse anti-TRF1 antibody (GeneTex; GTX70304). mouse anti-TRF2 antibody (Millipore; 05-521), rabbit anti-coilin antibody (GeneTex; GTX112570).

    Article Title: Recruitment of TRF2 to laser-induced DNA damage sites.
    Article Snippet: Several lines of evidence suggest that the telomere-associated protein TRF2 plays critical roles in the DNA damage response.. TRF2 is rapidly and transiently phosphorylated by an ATM-dependent pathway in response to DNA damage and this DNA damage-induced phosphoryation is essential for the DNA-PKdependent pathway of DNA double-strand break repair (DSB).. However, the type of DNA damage that induces TRF2 localization to the damage sites, the requirement for DNA damage-induced phosphorylation of TRF2 for its recruitment, as well as the detailed kinetics of TRF2 accumulation at DNA damage sites have not been fully investigated.

    Article Title: MiR-182-3p targets TRF2 and impairs tumor growth of triple-negative breast cancer.
    Article Snippet: The telomeric repeat-binding factor 2 (TRF2) is a telomere-capping protein that plays a key role in the maintenance of telomere structure and function.. It is highly expressed in different cancer types, and it contributes to cancer progression.. To date, anti-cancer strategies to target TRF2 remain a challenge.

    Two Tailed Test:

    Article Title: MiR ‐182‐3p targets TRF2 and impairs tumor growth of triple‐negative breast cancer
    Article Snippet: For immune labeling, cells were incubated with mouse anti-TRF2 (1:2,000, Millipore, 4A794) antibody in 3% BSA, 0.1% Tween-20 (1× PBS) for 1 h at RT.

    Article Title: The structurally conserved TELR region on shelterin protein TPP1 is essential for telomerase processivity but not recruitment
    Article Snippet: Immunofluorescence staining and FISH were carried out as described previously ( ) with the following antibodies: rabbit anti-GFP antibody (Novus; NB600-308), mouse anti-TRF1 antibody (GeneTex; GTX70304). mouse anti-TRF2 antibody (Millipore; 05-521), rabbit anti-coilin antibody (GeneTex; GTX112570).

    Article Title: Recruitment of TRF2 to laser-induced DNA damage sites.
    Article Snippet: Several lines of evidence suggest that the telomere-associated protein TRF2 plays critical roles in the DNA damage response.. TRF2 is rapidly and transiently phosphorylated by an ATM-dependent pathway in response to DNA damage and this DNA damage-induced phosphoryation is essential for the DNA-PKdependent pathway of DNA double-strand break repair (DSB).. However, the type of DNA damage that induces TRF2 localization to the damage sites, the requirement for DNA damage-induced phosphorylation of TRF2 for its recruitment, as well as the detailed kinetics of TRF2 accumulation at DNA damage sites have not been fully investigated.

    Article Title: MiR-182-3p targets TRF2 and impairs tumor growth of triple-negative breast cancer.
    Article Snippet: The telomeric repeat-binding factor 2 (TRF2) is a telomere-capping protein that plays a key role in the maintenance of telomere structure and function.. It is highly expressed in different cancer types, and it contributes to cancer progression.. To date, anti-cancer strategies to target TRF2 remain a challenge.



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    a The diagram shows the workflow on a timeline: on the day after cell seeding, cells were treated with RHPS4 (0.2 and/or 0.5 μM). At 48 h after treatment, cells were again treated with RHPS4 (0.2 and/or 0.5 μM) or harvested and counted. At 72 h after the first treatment, the cell viability was checked, and finally at 96 h after the first treatment, the cells were harvested and processed. b Representative images showing U251MG cells stained by immunofluorescence using anti-BLM (red signals), and anti-TRF1 <t>or</t> <t>anti-TRF2</t> (green signals) antibodies. Merged images allow visualization of colocalizing dots (yellow signals). Yellow arrows indicate BLM and TRF1-TRF2 colocalizations. c Quantification of the colocalizations between BLM and TRF1 or TRF2 proteins in U251MG cell line upon RHPS4 treatment (0.2 and 0.5 μM). d , e Sensitivity of U251MG and BLM −/− cell lines to RHPS4 concentrations ranging from 0.01 to 2 μM (0.01; 0.125; 0.25; 0.5; 1; 2 μM), evaluated 96 h after the first treatment. Mitomycin C (MMC) was used as a positive control at concentrations of 0.1; 0.5; 1; 2; 5 μg/ml. The Sulforhodamine B (SRB) cytotoxicity assay showed that RHPS4 sensitivity was unchanged in U251MG and BLM −/− (IC50 was 0.56 μM in both cell lines). f Short-term cell proliferation in untreated cells, U251MG and BLM −/− , and in RHPS4-treated cells (0.5 μM) as evaluated 48 and 96 h after the first treatment. g Long-term cell proliferation in U251MG and BLM −/− untreated and RHPS4-treated cells (0.5 μM). Scale bars represent 5 μm. * p < 0.05, ** p < 0.01 (two-way ANOVA; n = 3). Error bars denote the standard deviation of the mean.
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    FAM111B -deficient cells have shorter telomeres. (A) Widefield images of chromosome spreads and (B) interphase cells from wild-type (WT) and FAM111B knockout cells stained with fluorescent probe against telomeric repeats. Scale bar 20 μm. (C) Quantification of individual TeloFISH foci intensity in metaphase spreads and interphase cells. Green bars show averages of N = 3, with at least 30 metaphase spreads or 60 cells scored per sample in each experiment. (D) Widefield images of wild-type (WT) and FAM111B −/- U2OS cells stained with <t>TRF2</t> antibodies. Scale bar 20 μm. (E) Quantification of TRF2 foci intensity in U2OS wild-type (WT) FAM111B negative U2OS cells. Green bars show averages of N = 3 experiments, where 400 cells was scored per sample in each experiment. (F) Quantification of TRF2 intensities in U2OS cells over-expressing empty vector (EV) or FLAG-tagged FAM111B variants wild-type (WT), protease-dead (PD) or HFP mutant (Q430P, QP), respectively. Green bars show averages of N = 3 experiments, with at least 50 cells per sample scored in each experiment. K-W test was used in C, E and (F) .
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    FAM111B -deficient cells have shorter telomeres. (A) Widefield images of chromosome spreads and (B) interphase cells from wild-type (WT) and FAM111B knockout cells stained with fluorescent probe against telomeric repeats. Scale bar 20 μm. (C) Quantification of individual TeloFISH foci intensity in metaphase spreads and interphase cells. Green bars show averages of N = 3, with at least 30 metaphase spreads or 60 cells scored per sample in each experiment. (D) Widefield images of wild-type (WT) and FAM111B −/- U2OS cells stained with <t>TRF2</t> antibodies. Scale bar 20 μm. (E) Quantification of TRF2 foci intensity in U2OS wild-type (WT) FAM111B negative U2OS cells. Green bars show averages of N = 3 experiments, where 400 cells was scored per sample in each experiment. (F) Quantification of TRF2 intensities in U2OS cells over-expressing empty vector (EV) or FLAG-tagged FAM111B variants wild-type (WT), protease-dead (PD) or HFP mutant (Q430P, QP), respectively. Green bars show averages of N = 3 experiments, with at least 50 cells per sample scored in each experiment. K-W test was used in C, E and (F) .
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    Image Search Results


    a The diagram shows the workflow on a timeline: on the day after cell seeding, cells were treated with RHPS4 (0.2 and/or 0.5 μM). At 48 h after treatment, cells were again treated with RHPS4 (0.2 and/or 0.5 μM) or harvested and counted. At 72 h after the first treatment, the cell viability was checked, and finally at 96 h after the first treatment, the cells were harvested and processed. b Representative images showing U251MG cells stained by immunofluorescence using anti-BLM (red signals), and anti-TRF1 or anti-TRF2 (green signals) antibodies. Merged images allow visualization of colocalizing dots (yellow signals). Yellow arrows indicate BLM and TRF1-TRF2 colocalizations. c Quantification of the colocalizations between BLM and TRF1 or TRF2 proteins in U251MG cell line upon RHPS4 treatment (0.2 and 0.5 μM). d , e Sensitivity of U251MG and BLM −/− cell lines to RHPS4 concentrations ranging from 0.01 to 2 μM (0.01; 0.125; 0.25; 0.5; 1; 2 μM), evaluated 96 h after the first treatment. Mitomycin C (MMC) was used as a positive control at concentrations of 0.1; 0.5; 1; 2; 5 μg/ml. The Sulforhodamine B (SRB) cytotoxicity assay showed that RHPS4 sensitivity was unchanged in U251MG and BLM −/− (IC50 was 0.56 μM in both cell lines). f Short-term cell proliferation in untreated cells, U251MG and BLM −/− , and in RHPS4-treated cells (0.5 μM) as evaluated 48 and 96 h after the first treatment. g Long-term cell proliferation in U251MG and BLM −/− untreated and RHPS4-treated cells (0.5 μM). Scale bars represent 5 μm. * p < 0.05, ** p < 0.01 (two-way ANOVA; n = 3). Error bars denote the standard deviation of the mean.

    Journal: Communications Biology

    Article Title: BLM and FANCJ role in the response to G-quadruplex-dependent telomeric replicative stress

    doi: 10.1038/s42003-025-09367-z

    Figure Lengend Snippet: a The diagram shows the workflow on a timeline: on the day after cell seeding, cells were treated with RHPS4 (0.2 and/or 0.5 μM). At 48 h after treatment, cells were again treated with RHPS4 (0.2 and/or 0.5 μM) or harvested and counted. At 72 h after the first treatment, the cell viability was checked, and finally at 96 h after the first treatment, the cells were harvested and processed. b Representative images showing U251MG cells stained by immunofluorescence using anti-BLM (red signals), and anti-TRF1 or anti-TRF2 (green signals) antibodies. Merged images allow visualization of colocalizing dots (yellow signals). Yellow arrows indicate BLM and TRF1-TRF2 colocalizations. c Quantification of the colocalizations between BLM and TRF1 or TRF2 proteins in U251MG cell line upon RHPS4 treatment (0.2 and 0.5 μM). d , e Sensitivity of U251MG and BLM −/− cell lines to RHPS4 concentrations ranging from 0.01 to 2 μM (0.01; 0.125; 0.25; 0.5; 1; 2 μM), evaluated 96 h after the first treatment. Mitomycin C (MMC) was used as a positive control at concentrations of 0.1; 0.5; 1; 2; 5 μg/ml. The Sulforhodamine B (SRB) cytotoxicity assay showed that RHPS4 sensitivity was unchanged in U251MG and BLM −/− (IC50 was 0.56 μM in both cell lines). f Short-term cell proliferation in untreated cells, U251MG and BLM −/− , and in RHPS4-treated cells (0.5 μM) as evaluated 48 and 96 h after the first treatment. g Long-term cell proliferation in U251MG and BLM −/− untreated and RHPS4-treated cells (0.5 μM). Scale bars represent 5 μm. * p < 0.05, ** p < 0.01 (two-way ANOVA; n = 3). Error bars denote the standard deviation of the mean.

    Article Snippet: The slides were then incubated in blocking buffer (1% BSA dissolved in 1X PBS [w/v]) and then incubated overnight (ON) at 4 °C with the primary antibodies (rabbit polyclonal anti-BLM (#A300-110A, Bethyl) (1:100); mouse monoclonal anti-TRF1 (4E4 clone, GTX70304, GeneTex) (1:20) or mouse monoclonal anti-TRF2 (9F10 clone, sc-47693, Santa Cruz Biotechnology) (1:100)).

    Techniques: Staining, Immunofluorescence, Positive Control, Cytotoxicity Assay, Standard Deviation

    FAM111B -deficient cells have shorter telomeres. (A) Widefield images of chromosome spreads and (B) interphase cells from wild-type (WT) and FAM111B knockout cells stained with fluorescent probe against telomeric repeats. Scale bar 20 μm. (C) Quantification of individual TeloFISH foci intensity in metaphase spreads and interphase cells. Green bars show averages of N = 3, with at least 30 metaphase spreads or 60 cells scored per sample in each experiment. (D) Widefield images of wild-type (WT) and FAM111B −/- U2OS cells stained with TRF2 antibodies. Scale bar 20 μm. (E) Quantification of TRF2 foci intensity in U2OS wild-type (WT) FAM111B negative U2OS cells. Green bars show averages of N = 3 experiments, where 400 cells was scored per sample in each experiment. (F) Quantification of TRF2 intensities in U2OS cells over-expressing empty vector (EV) or FLAG-tagged FAM111B variants wild-type (WT), protease-dead (PD) or HFP mutant (Q430P, QP), respectively. Green bars show averages of N = 3 experiments, with at least 50 cells per sample scored in each experiment. K-W test was used in C, E and (F) .

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Loss of FAM111B protease mutated in hereditary fibrosing poikiloderma negatively regulates telomere length

    doi: 10.3389/fcell.2023.1175069

    Figure Lengend Snippet: FAM111B -deficient cells have shorter telomeres. (A) Widefield images of chromosome spreads and (B) interphase cells from wild-type (WT) and FAM111B knockout cells stained with fluorescent probe against telomeric repeats. Scale bar 20 μm. (C) Quantification of individual TeloFISH foci intensity in metaphase spreads and interphase cells. Green bars show averages of N = 3, with at least 30 metaphase spreads or 60 cells scored per sample in each experiment. (D) Widefield images of wild-type (WT) and FAM111B −/- U2OS cells stained with TRF2 antibodies. Scale bar 20 μm. (E) Quantification of TRF2 foci intensity in U2OS wild-type (WT) FAM111B negative U2OS cells. Green bars show averages of N = 3 experiments, where 400 cells was scored per sample in each experiment. (F) Quantification of TRF2 intensities in U2OS cells over-expressing empty vector (EV) or FLAG-tagged FAM111B variants wild-type (WT), protease-dead (PD) or HFP mutant (Q430P, QP), respectively. Green bars show averages of N = 3 experiments, with at least 50 cells per sample scored in each experiment. K-W test was used in C, E and (F) .

    Article Snippet: Primary antibodies were as follows: mouse anti-human BLM (sc-365753, Santa Cruz Biotechnology) at 1:250 (IF), mouse anti-BrdU (347580, Becton Dickinson) at 1:20 (Flow cytometry), rabbit anti-human FAM111B (HPA038637, Atlas Antibodies) at 1:1,000 (IB and IF), mouse anti-human Histone H3 (10799, Abcam) at 1:1,000 (IB), mouse anti-human Lamin A/C (sc-376248, Santa Cruz Biotechnology) at 1:1,000 (IB and IF), rabbit anti-human Lamin B1 (12987-1-AP, Proteintech Group) at 1:1,000 (IB and IF), rabbit anti-human NUP42 (16587-1-AP, Proteintech Group) at 1:1,000 (IB), mouse anti-human PCNA (sc-56, Santa Cruz Biotechnology) at 1:1,000 (IF), rabbit anti-human SEC13 (15397-1-AP, Proteintech Group) at 1:1,000 (IB), mouse anti-human TRF2 (NB100-56506, Novus Biologicals) at 1:1,000 (IF), mouse anti-human αTubulin (T6074, Sigma) at 1:10,000 (WB) or 1:1,000 (IF).

    Techniques: Knock-Out, Staining, Expressing, Plasmid Preparation, Mutagenesis