rabbit anti ctip polyclonal antibody  (Bethyl)

Journal: Nature Communications

Article Title: DHX9-dependent recruitment of BRCA1 to RNA promotes DNA end resection in homologous recombination

doi: 10.1038/s41467-021-24341-z

Figure Lengend Snippet: a Fluorescent image (left panel) and quantification (right panel) showing the decreased formation of RAD51 foci in control U2OS cells (red) and cells knocked down for DHX9 (blue) treated with 1 μM camptothecin for 2 h then left for 2 h to recover. b DHX9 is in a common genetic pathway for the formation of RPA foci with CTIP and MRE11. RPA foci in cells knocked down using siRNA for the indicated genes are shown. Cells were treated with 1 μM camptothecin for 2 h. c Recruitment of DHX9 to foci is dependent on ATM and ATR. Wild-type cells were inhibited for ATR and ATM using VE-821 and KU55933, respectively. d , e DHX9 is in a common genetic pathway for the formation of RPA foci with ATR and ATM. RPA foci are shown for cells treated with inhibitors for ATR (VE-821) and ATM (KU55933) ( d ) or knocked down with siRNA against ATM and ATR ( e ). f Western blot showing that knockdown of DHX9 impairs Cpt induced autophosphorylation of ATR on Thr1989 and phosphorylation of Chk1 on ser 345. Quantification of n cells (as indicated) from three pooled biologically independent experiments were performed in ( a – e ). Means of data sets were shown to be significantly different using one-way ANOVA with Tukey’s post hoc test. (ns not significant, * p < 0.1, ** p < 0.01, *** p < 0.001, and **** p < 0.0001). Error bars indicating one standard deviation are also indicated. Source data are provided as a Source Data file.

Article Snippet: Rabbit polyclonal anti-RNA Helicase A (ab26271, Abcam, for IP 5 μg/ml, IF 1:1000, and WB 1:2000 dilution), mouse monoclonal anti-BRCA1 antibody (OP92, Ab-1; Calbiochem, for IPs 10 μg/ml, IF 1:250, and WB 1:200 dilution), mouse anti-RPA32 (RPA2 Ab #2; Calbiochem, IF 1:500, WB 1:1000, and for FACS 1:100 dilution), rabbit polyclonal anti-RNA-Polymerase II (N-20; Santa Cruz Biotechnology, IP 4 μg/ml and WB 1:1000 dilution), mouse monoclonal anti-GAPDH antibody (GT239; GeneTex, WB 1:1000 dilution), mouse monoclonal anti-phospho-Histone H2A.X (clone JBW301; EMD Millipore, IF 1:250 dilution and WB 1:500 dilution), rabbit anti-phospho-Histone H2A.X (ab81299, Abcam, IF 1:1000 dilution), rabbit polyclonal anti-Rad51 antibody (H-92; Santa Cruz Biotechnology, IF 1:250 dilution), rabbit polyclonal anti-53BP1 Antibody (NB100-904; Novus Biologicals, IF 1:500, WB 1:2000), rabbit polyclonal anti-53BP1 Antibody (A300-272A, Universal biologicals, IF 1:1000 dilution), mouse monoclonal Anti-BrdU antibody (B44; BD Biosciences, 1:1000 dilution for IF), Rat monoclonal anti-BrdU antibody (ab6326, Abcam, IF 1:1000 dilution), rabbit polyclonal anti-CTIP antibody (NB100-79810, Novus Biologicals, WB 1:2000, IF 1:500 dilution), rabbit polyclonal anti-MRE 11 antibody (NB100-142, Novus Biologicals, WB 1:1000 dilution), rabbit polyclonal anti-BLM antibody, (PLA0029, Sigma-Aldrich, WB 1:1000 dilution), mouse monoclonal anti-ATR antibody (NB100-308, Novus Biologicals,WB 1:1000 dilution), rabbit polyclonal anti-ATM antibody (NB100-104, Novus Biologicals, WB 1:1000 dilution), rabbit monoclonal anti-Ku80 Antibody (218, Cell Signaling, WB 1:1000 dilution).

Techniques: Control, Western Blot, Knockdown, Phospho-proteomics, Standard Deviation

Journal: Nature Communications

Article Title: DHX9-dependent recruitment of BRCA1 to RNA promotes DNA end resection in homologous recombination

doi: 10.1038/s41467-021-24341-z

Figure Lengend Snippet: a Fluorescence images (left panel) and graph (right panel) showing that localization of BLM to camptothecin-induced DNA damage foci is impaired in cells knocked down for DHX9. b Fluorescence images (left panel) and graph (right panel) showing that localization of CTIP to camptothecin-induced DNA damage foci is impaired in cells knocked down for DHX9. Quantification of n cells (as indicated) from three pooled biologically independent experiments were performed in ( a ) and ( b ). Means were shown to be significantly different using one-way ANOVA with post hoc Tukey’s test (**** p < 0.0001). c Western blot of fractionated cell extracts showing that localization of BLM and CTIP to chromatin (P1 fraction) in response to camptothecin-induced DNA damage is reduced in cells knocked down for DHX9. Localization of BLM and CTIP in cytoplasmic (S1) and nuclear fractions (S2) is not decreased. Histone H3 is shown as a marker of S2 and P1 fractions. d DNA synthesis is impaired in DHX9 and BRCA1 deficient cells treated with camptothecin (5 μM for 2 h). This defect is not suppressed by knockdown of 53BP1 Right panel shows representative images for the incorporation of CldU and IdU nucleotide analogs as well as merged images. The left panel shows graphical data of cells stained with both CldU and IldU as a percentage of total cells stained with CldU. Graphs include data from three biologically independent experiments. Mean and error bars indicating one standard deviation are also indicated. Statistical significance for all experiments was demonstrated using one-way ANOVA with post hoc Tukey’s test (**** p < 0.0001, * p < 0.1, ns not significant). Source data are provided as a Source Data file.

Article Snippet: Rabbit polyclonal anti-RNA Helicase A (ab26271, Abcam, for IP 5 μg/ml, IF 1:1000, and WB 1:2000 dilution), mouse monoclonal anti-BRCA1 antibody (OP92, Ab-1; Calbiochem, for IPs 10 μg/ml, IF 1:250, and WB 1:200 dilution), mouse anti-RPA32 (RPA2 Ab #2; Calbiochem, IF 1:500, WB 1:1000, and for FACS 1:100 dilution), rabbit polyclonal anti-RNA-Polymerase II (N-20; Santa Cruz Biotechnology, IP 4 μg/ml and WB 1:1000 dilution), mouse monoclonal anti-GAPDH antibody (GT239; GeneTex, WB 1:1000 dilution), mouse monoclonal anti-phospho-Histone H2A.X (clone JBW301; EMD Millipore, IF 1:250 dilution and WB 1:500 dilution), rabbit anti-phospho-Histone H2A.X (ab81299, Abcam, IF 1:1000 dilution), rabbit polyclonal anti-Rad51 antibody (H-92; Santa Cruz Biotechnology, IF 1:250 dilution), rabbit polyclonal anti-53BP1 Antibody (NB100-904; Novus Biologicals, IF 1:500, WB 1:2000), rabbit polyclonal anti-53BP1 Antibody (A300-272A, Universal biologicals, IF 1:1000 dilution), mouse monoclonal Anti-BrdU antibody (B44; BD Biosciences, 1:1000 dilution for IF), Rat monoclonal anti-BrdU antibody (ab6326, Abcam, IF 1:1000 dilution), rabbit polyclonal anti-CTIP antibody (NB100-79810, Novus Biologicals, WB 1:2000, IF 1:500 dilution), rabbit polyclonal anti-MRE 11 antibody (NB100-142, Novus Biologicals, WB 1:1000 dilution), rabbit polyclonal anti-BLM antibody, (PLA0029, Sigma-Aldrich, WB 1:1000 dilution), mouse monoclonal anti-ATR antibody (NB100-308, Novus Biologicals,WB 1:1000 dilution), rabbit polyclonal anti-ATM antibody (NB100-104, Novus Biologicals, WB 1:1000 dilution), rabbit monoclonal anti-Ku80 Antibody (218, Cell Signaling, WB 1:1000 dilution).

Techniques: Fluorescence, Western Blot, Marker, DNA Synthesis, Knockdown, Staining, Standard Deviation

Journal: Cell reports

Article Title: Dual Processing of R-Loops and Topoisomerase I Induces Transcription-Dependent DNA Double-Strand Breaks

doi: 10.1016/j.celrep.2019.08.041

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: CtIP (RBBP8) rabbit polyclonal antibody (used for: WB) , Bethyl , Cat# A300-488A; RRID:AB_2175262.

Techniques: Recombinant, Transfection, Viability Assay, Imaging, Luciferase, Control, Software, Fluorescence, Microscopy