rabbit anti setx (Bethyl)
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Bethyl
rabbit anti setx
Rabbit Anti Setx, supplied by Bethyl, used in various techniques. Bioz Stars score: 93/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+setx/Senataxin+Antibody/bio_rxiv__2025__09__23__678005-123-26-29
Average 93 stars, based on 26 article reviews
Rabbit Anti Setx, supplied by Bethyl, used in various techniques. Bioz Stars score: 93/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+setx/Senataxin+Antibody/bio_rxiv__2025__09__23__678005-123-26-29
Average 93 stars, based on 26 article reviews
rabbit anti setx - by Bioz Stars,
2026-09
93/100 stars
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Western Blot:Article Title: PARP1 associates with R-loops to promote their resolution and genome stability Article Snippet: The next day, the media was replaced with standard growth media and the cells were allowed to recover for 24 h. Knockdown efficiency for RNH1 siRNA was validated by western blot on a standard SDS-PAGE gel using the anti-RNase H1 (H-4, Santa Cruz Biotechnology #sc-376326) and anti-β-actin antibodies. .. To validate SETX siRNA efficiency, western blot was performed with a NuPAGE 3–8% Tris-acetate gel (ThermoFisher Scientific) using the Article Title: PARP1 associates with R-loops to promote their resolution and genome stability. Article Snippet: The next day, the media was replaced with standard growth media and the cells were allowed to recover for 24 h. Knockdown efficiency for RNH1 siRNA was validated by western blot on a standard SDS-PAGE gel using the anti-RNase H1 (H-4, Santa Cruz Biotechnology #sc-376326) and anti-actin antibodies. .. To validate SETX siRNA efficiency, western blot was performed with a NuPAGE 3–8% Trisacetate gel (ThermoFisher Scientific) using the Article Title: SUGP1 loss is the sole driver of SF3B1 hotspot mutant missplicing in cancer Article Snippet: .. Western blotting was performed as described with the following primary antibodies: anti-AQR (1:1,000, Bethyl Laboratories, A302-547A-T), anti-SUGP1 (1:1,000, Sigma-Aldrich, HPA004890), anti-ACTIN (1:2,000, Sigma-Aldrich, A2066), Article Title: SUGP1 loss drives SF3B1 hotspot mutant missplicing in cancer Article Snippet: .. Western blotting was performed as described with the following primary antibodies: anti-AQR (1:1,000, Bethyl Laboratories, A302-547A-T), anti-SUGP1 (1:1,000, Sigma-Aldrich, HPA004890), anti-ACTIN (1:2,000, Sigma-Aldrich, A2066), other:Article Title: SUGP1 loss drives SF3B1 hotspot mutant missplicing in cancer Article Snippet: anti-SETX , Bethyl Laboratories , A301-105A. Incubation:Article Title: RNF8 ubiquitylation of XRN2 facilitates R-loop resolution and restrains genomic instability in BRCA1 mutant cells Article Snippet: .. For each IP, 50 μl of chromatin was diluted 10 times in IP dilution buffer (0.01% SDS, 1.1% TritonX-100, 1.1 mM EDTA, 20 mM Tris–HCl pH 8.0, 167 nM NaCl) and incubated with 3 μg anti-XRN2 (Bethyl, A301-103A), 4 μg Article Title: RNF8 ubiquitylation of XRN2 facilitates R-loop resolution and restrains genomic instability in BRCA1 mutant cells. Article Snippet: .. For each IP, 50 l of chromatin was diluted 10 times in IP dilution buffer (0.01% SDS, 1.1% TritonX-100, 1.1 mM EDTA, 20 mM Tris–HCl pH 8.0, 167 nM NaCl) and incubated with 3 g anti-XRN2 (Bethyl, A301-103A), 4 g Control:Article Title: RNF8 ubiquitylation of XRN2 facilitates R-loop resolution and restrains genomic instability in BRCA1 mutant cells Article Snippet: .. For each IP, 50 μl of chromatin was diluted 10 times in IP dilution buffer (0.01% SDS, 1.1% TritonX-100, 1.1 mM EDTA, 20 mM Tris–HCl pH 8.0, 167 nM NaCl) and incubated with 3 μg anti-XRN2 (Bethyl, A301-103A), 4 μg Article Title: RNF8 ubiquitylation of XRN2 facilitates R-loop resolution and restrains genomic instability in BRCA1 mutant cells. Article Snippet: .. For each IP, 50 l of chromatin was diluted 10 times in IP dilution buffer (0.01% SDS, 1.1% TritonX-100, 1.1 mM EDTA, 20 mM Tris–HCl pH 8.0, 167 nM NaCl) and incubated with 3 g anti-XRN2 (Bethyl, A301-103A), 4 g SDS Page:Article Title: Dual Processing of R-Loops and Topoisomerase I Induces Transcription-Dependent DNA Double-Strand Breaks Article Snippet: .. Proteins were separated by SDS-PAGE and immunoblotted with the following antibodies at dilutions recommended by the manufacturer: anti-pan-actin (MAB1501; Millipore), anti-ARTEMIS (#13381; Cell Signaling Technology), antimCherry (GTX128509; GeneTex), anti-CtIP (A300–488A; Bethyl), anti-FEN1 (A300–255A; Bethyl), anti-KAP1 (A300–274A; Bethyl), anti-DNA ligase3 (A301–636A; Bethyl), anti-MRE11 (A303–998A; Bethyl), anti-MUS81 (ab14387; Abcam), anti-PNKP (A300–257A; Bethyl), |
![<t>SETX-depleted</t> cells exhibit R-loop-mediated replication stress. ( A ) Western blot analysis to check SETX expression levels and CHK1-S345 phosphorylation in U2OS cells transfected with two different siRNAs against SETX for 72 h. ( B ) Western blot analysis to confirm over-expression of RNaseH1-GFP in U2OS T-REx [RNaseH1-GFP] cells treated with doxycycline (Dox; 1 ng/ml) for 24 h. ( C ) Schematic of DNA fiber labeling and quantification of replication tract lengths in mock (siLUC)- and SETX-depleted U2OS T-REx [RNaseH1-GFP] cells. 1 ng/ml Dox was added 24 h before the labeling to induce RNase H1-GFP over-expression. Dimethylsulphoxide (DMSO) was used as a vehicle. The values of CldU + IdU tract lengths measured in three independent experiments are plotted (n > 300). Red lines represent the median. ( D ) Quantification of replication fork stalling events on DNA fibers in ( C ). Replication tracts containing only the first label (CldU-only tracts) were designated as stalled replication forks while the replication tracts containing both labels (CldU + IdU) were scored as ongoing forks. Data represent mean ± SD ( n = 3). ( E ) Representative images and quantification of RNaseH1(D210N)-GFP foci in U2OS T-REx [RNaseH1(D210N)-GFP] cells transfected with indicated siRNAs. PCNA immunofluorescence staining was used to identify S-phase cells (PCNA + ). ( F ) Representative images and quantification of PLA foci between PCNA and elongating form of RNA polymerase II (RNAPIIS2P) in U2OS cells transfected with indicated siRNAs. EdU incorporation was performed to mark S-phase cells. Triptolide (1 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\mu$\end{document} M) was added 2 h prior to PLA. (E, F) Medians of data sets from three independent experiments are plotted (n > 200). Data are presented as mean ± SD. Scale bar, 10 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\mu {\mathrm{m}}$\end{document} . Statistical analysis: Kruskal–Wallis test followed by Dunn's multiple comparisons test was used in ( C ). Ordinary one-way ANOVA followed by Šídák's multiple comparisons, with a single pooled variance was used in (D)–(F).](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_7401/pmc11417401/pmc11417401__gkae673fig1.jpg)
![SETX depletion induces R-loop-dependent mitotic DNA synthesis and micronucleation. ( A ) Schematic representation of MiDAS assay, representative images and quantification of EdU incorporation events on metaphase chromosome spreads of U2OS T-REx [RNaseH1-GFP] cells transfected with indicated siRNAs. Cells were treated with 9 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\mu$\end{document} M CDK1 inhibitor RO-3306 for 16 h, and then released into mitosis in the presence of 20 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\mu$\end{document} M EdU and colcemid. Where indicated, over-expression of RNaseH1 was induced by the addition of doxycycline (Dox; 1 ng/ml) for 24 h. Dimethylsulphoxide (DMSO) was used as a vehicle. Data represent mean ± SD ( n = 3). At least 50 metaphase spreads were analyzed for the presence of EdU foci in each experiment. Scale bar, 10 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\mu {\mathrm{m}}.$\end{document} ( B ) Experimental workflow with a representative image and quantification of micronuclei in U2OS T-REx [RNaseH1-GFP] cells transfected with indicated siRNAs. Cytochalasin B (2 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\mu$\end{document} g/ml) was added 16 h prior to fixation to arrest cells before cytokinesis. Dox (1 ng/ml) was added to cells to induce RNaseH1-GFP over-expression. Data represent mean ± SD ( n = 4). At least 300 binucleated cells were scored in each experiment. Scale bar, 10 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\mu {\mathrm{m}}.$\end{document} Red arrow indicates micronucleus. ( C ) Experimental workflow with a representative image and quantification of 53BP1 nuclear bodies in U2OS T-REx [RNaseH1-GFP] cells transfected with indicated siRNAs. EdU incorporation was used to exclude S-phase cells from quantification. Dox (1 ng/ml) was added to cells to induce RNaseH1-GFP over-expression. Data represent mean ± SD ( n = 3). At least 300 EdU-negative cells were analyzed in each experiment. Scale bar, 10 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\mu {\mathrm{m}}.$\end{document} Statistical analysis: Ordinary one-way ANOVA followed by Šídák's multiple comparisons, with a single pooled variance was used in (A)–(C).](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_7401/pmc11417401/pmc11417401__gkae673fig2.jpg)
![SETX is essential for restarting R-loop-stalled forks via MUS81-LIG4-ELL axis. ( A ) Effect of SETX depletion on replication fork recovery after induction of R-loop-mediated fork stalling in U2OS cells. Schematic of DNA fiber assays is shown on the top. The R-loop-inducing drug, camptothecin (CPT; 100 nM) was added to cells during CldU incorporation for the last 20 min. Dimethylsulphoxide (DMSO) was used as a vehicle. Cells were then washed and released into a fresh medium containing IdU. Replication tracts containing only CldU (red-only tracts) were designated as stalled replication forks. Replication tracts containing both CldU and IdU were scored as ongoing forks (red/green tracts). The percentage of stalled replication forks is plotted. Data represent mean ± SD ( n = 3). ( B ) Effect of SETX depletion on the rescue of CPT-induced fork stalling by PARP inhibition in U2OS cells, which requires the MUS81-LIG4-ELL axis. Schematic of DNA fiber assays and representative images of DNA replication tracts upon indicated conditions are shown on the left. CPT (100 nM) was present during IdU labeling. The PARP inhibitor (PARPi), olaparib (10 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\mu$\end{document} M) was added 2 h before DNA fiber labeling and was also present during the labeling to boost replication restart. The values of the IdU/CldU tract length ratio obtained in three independent experiments are plotted ( n > 300). Red lines represent the median. Statistical analysis: Ordinary one-way ANOVA followed by Šídák's multiple comparisons, with a single pooled variance was used in (A). Kruskal–Wallis test followed by Dunn's multiple comparisons test was used in (B). Scale bar, 10 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\mu$\end{document} m.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_7401/pmc11417401/pmc11417401__gkae673fig3.jpg)
![SETX depletion induces MUS81-dependent nascent DNA degradation at R-loop-stalled forks. ( A ) Schematic of fork degradation assay. U2OS T-REx [RNaseH1-GFP] or U2OS cells were transfected with appropriate siRNA and after 72 h were subjected to DNA fiber labeling, followed by hydroxyurea (HU; 4 mM) treatment for 5 h to induce replication fork stalling. Representative images of replication tracts on DNA fibers of mock (siLUC)- and SETX-depleted (siSETX-1) U2OS cells are also shown. Scale bar, 10 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\mu$\end{document} m. ( B ) Quantification of HU-induced nascent DNA degradation in U2OS T-REx [RNaseH1-GFP] cells transfected with indicated siRNAs. Where indicated, doxycycline (Dox; 1 ng/ml) was added 24 h prior to labeling to induce RNaseH1-GFP over-expression, which eliminates R-loops. (C–F) Quantification of HU-induced nascent DNA degradation in U2OS cells transfected with indicated siRNAs. Where indicated in ( C ), transcription inhibitors, DRB (100 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\mu$\end{document} M) or triptolide (TRP, 1 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\mu$\end{document} M), were added 2 h prior to labeling and were present throughout the labeling and HU treatment. In ( D ), SETX or BRCA2 were co-depleted with either ZRANB3 or HLTF to prevent fork reversal. Bottom panel shows western blot to confirm the depletion of indicated proteins. In ( E ), SETX was co-depleted with MUS81 to prevent fork cleavage. Bottom panel shows western blot to confirm the depletion of proteins. Where indicated in ( F ), MRE11 inhibitor, Mirin (50 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\mu$\end{document} M) and the DNA2 inhibitor, C5 (25 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} $\mu$\end{document} M) were added along with HU. In (B)–(F), the values of the IdU/CldU tract length ratio obtained in three independent experiments are plotted ( n > 300). Red lines represent the median. Statistical analysis: Kruskal–Wallis test followed by Dunn's multiple comparisons test was used in (B)–(F).](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_7401/pmc11417401/pmc11417401__gkae673fig4.jpg)
![SETX forms a complex with DDX17 in human cells. ( A ) DDX17 isoforms (p72 and p82) co-immunoprecipitate with SETX from HEK293 cell extracts. Cells were treated either with 100 nM CPT or with vehicle (DMSO) for 1 h before harvest. Cell extracts were immunoprecipitated with rabbit polyclonal anti-SETX antibody. Rabbit IgG was incubated with extract of DMSO-treated cells as a control. Representative western blot of three independent experiments is shown. ( B ) Representative images (left panel) and quantification (right panel) of PLA foci between SETX and DDX17 in U2OS T-REx [RNaseH1-GFP] cells before and 24 h after induction of RNaseH1 expression with doxycycline (Dox). Medians of data sets from three independent experiments are plotted (n > 200). Data are presented as mean ± SD. Statistical analysis: Ordinary one-way ANOVA followed by Šídák's multiple comparisons, with a single pooled variance. Scale bar, 10 μ m.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_7401/pmc11417401/pmc11417401__gkae673fig7.jpg)
