rabbit anti orc2  (Bethyl)

Journal: EMBO Reports

Article Title: The nucleolar protein GNL3 prevents resection of stalled replication forks

doi: 10.15252/embr.202357585

Figure Lengend Snippet: Scheme explaining the mini esiRNA screen to determine the best candidates for further characterization. HCT116 cells were transfected in 96‐wells plate with each esiRNA from the library (Dataset ). After 48 h cells were treated with 1 μM camptothecin for 4 h and subjected to immunofluorescence using an antibody directed against γH2A.X. The level of γH2A.X within nuclei was analyzed using a Celigo high‐throughput microscope. The γH2A.X level upon depletion and camptothecin treatment in five biological replicates was used to rank the candidates, GNL3 was ranked first and EGFP (negative control) was ranked at the end of the list (21 st ). The bounds of the box are the 25th and 75th percentiles, the line in the center is the median and the bounds of the whiskers are the maxima and the minima. Graphic representation (related to Fig ) of the medians of CldU/IdU ratios in three biological replicates, the average is indicated in red. Western‐blot analysis of HeLa S3 cells treated with 1 μM camptothecin (CPT) during the indicated time. Western‐blot analysis of HeLa S3 cells treated with 10 μM etoposide (ETP) during the indicated time. Graphic representation (related to Fig ) of the medians of CldU/IdU ratios in three biological replicates, the average is indicated in red. HeLa S3 cells were sequentially labeled for 30 min with IdU and for 30 min with CldU then treated with 1 μM CPT for 240 min. The ratio between CldU and IdU is plotted, and the red line indicates the median. For statistical analysis, Mann–Whitney test was used; **** P < 0.0001. At least 100 individual DNA fibers was counted for each condition, the graphic representation of the medians of CldU/IdU ratios in three biological replicates with the average indicated in red is shown. HeLa S3 cells were sequentially labeled for 30 min with IdU and for 30 min with CldU then treated with 10 μM ETP for 120 min. The ratio between CldU and IdU is plotted, and the red line indicates the median. For statistical analysis, Mann–Whitney test was used; **** P < 0.0001. At least 100 individual DNA fibers was counted for each condition, the graphic representation of the medians of CldU/IdU ratios in three biological replicates with the average indicated in red is shown. Western‐blot analysis of HeLa S3 cells depleted for GNL3, MRE11, CtIP or EXO1. Graphic representation (related to Fig ) of the medians of CldU/IdU ratios in three biological replicates, the average is indicated in red. Western‐blot analysis of Flp‐in T‐Rex HeLa cells expressing GNL3‐WT tagged with FLAG. Cells were first transfected with siControl or siGNL3 for 48 h then expression of GNL3‐WT (resistant to the siRNA against GNL3) was induced using 10 μg/ml of doxycycline (DOX) for 16 h. Graphic representation (related to Fig ) of the medians of CldU/IdU ratios in three biological replicates, the average is indicated in red. Western‐blot analysis of the indicated proteins upon chromatin fractionation of Hela S3 cells treated 4 h with 5 mM HU. Source data are available online for this figure.

Article Snippet: Immunoprecipitations were performed overnight at 4°C with protein G Dynabeads (Thermo Fisher Scientific) coupled to either rabbit immunoglobulin G (IgG) (P120‐201, Bethyl Laboratories), rabbit GNL3 (sc‐166460, Santa Cruz Biotechnology) or rabbit ORC2 antibody (A302‐734A, Bethyl Laboratories).

Techniques: esiRNA, Transfection, Immunofluorescence, High Throughput Screening Assay, Microscopy, Negative Control, Western Blot, Labeling, MANN-WHITNEY, Expressing, Fractionation

Journal: EMBO Reports

Article Title: The nucleolar protein GNL3 prevents resection of stalled replication forks

doi: 10.15252/embr.202357585

Figure Lengend Snippet: Western‐blot analysis of HeLa S3 cells depleted with a pool of four siRNA‐targeting GNL3 (siGNL3) or not (siControl). HeLa S3 cells were sequentially labeled for 30 min with IdU and for 30 min with CldU with or without 1 μM CPT. Ratios between CldU and IdU are plotted, and the red line indicates the median. For statistical analysis, Mann–Whitney test was used; **** P < 0.0001. ns, not significant. 100 individual DNA fibers were counted for each condition, and biological replicates are shown in Fig . Western‐blot analysis of HeLa S3 cells treated with 5 mM HU during the indicated time. HeLa cells were sequentially labeled for 30 min with IdU and for 30 min with CldU then treated with 5 mM HU for 240 min. The ratio between CldU and IdU is plotted, and the red line indicates the median. For statistical analysis, Mann–Whitney test was used; **** P < 0.0001. 100 individual DNA fibers were counted for each condition, and biological replicates are shown in Fig . HeLa S3 were sequentially labeled for 30 min with IdU and for 30 min with CldU then treated with 5 mM HU for 240 min. The ratio between CldU and IdU is plotted, and the red line indicates the median. For statistical analysis, Mann–Whitney test was used; **** P < 0.0001. At least 80 individual DNA fibers were counted for each condition, and biological replicates are shown in Fig . Flp‐in T‐Rex HeLa cells were sequentially labeled for 30 min with IdU and for 30 min with CldU then treated with 5 mM HU for 240 min. The ratio between CldU and IdU is plotted, and the red line indicates the median. For statistical analysis, Mann–Whitney test was used; **** P < 0.0001. 100 individual DNA fibers were counted for each condition, and biological replicates are shown in Fig . Experimental set‐up of iPOND experiment. iPOND experiment analyzed by Western‐blot. Cells were pulsed with 15 min EdU and chased for 2 h with 10 μM thymidine or 5 mM HU. In no click sample, biotin‐TEG azide was replaced by DMSO. Source data are available online for this figure.

Article Snippet: Immunoprecipitations were performed overnight at 4°C with protein G Dynabeads (Thermo Fisher Scientific) coupled to either rabbit immunoglobulin G (IgG) (P120‐201, Bethyl Laboratories), rabbit GNL3 (sc‐166460, Santa Cruz Biotechnology) or rabbit ORC2 antibody (A302‐734A, Bethyl Laboratories).

Techniques: Western Blot, Labeling, MANN-WHITNEY

Journal: EMBO Reports

Article Title: The nucleolar protein GNL3 prevents resection of stalled replication forks

doi: 10.15252/embr.202357585

Figure Lengend Snippet: Western‐blot analysis upon treatment with HU and inhibition of CDC7, WEE1, or ATR. Graphic representation (related to Fig ) of the medians of CldU/IdU ratios in three biological replicates, the average is indicated in red. Graphic representation (related to Fig ) of the medians of CldU/IdU ratios in three biological replicates, the average is indicated in red. Graphic representation (related to Fig ) of the medians of CldU/IdU ratios in three biological replicates, the average is indicated in red. Quantification of RPA phosphorylation (S4/8) upon GNL3 depletion and CDC7 inhibition. The error bars represent the standard deviation between three biological replicates. Western‐blot analysis of HeLa S3 cells depleted or not for BRCA1 upon treatment with CDC7 inhibitor. Graphic representation (related to Fig ) of the medians of CldU/IdU ratios in three biological replicates, the average is indicated in red. Graphic representation (related to Fig ) of the medians of CldU/IdU ratios in three biological replicates, the average is indicated in red. Western‐blot analysis of HeLa S3 cells. Graphic representation (related to Fig ) of the medians of CldU/IdU ratios in three biological replicates, the average is indicated in red. Western‐blot analysis of control U‐2 OS cells (U‐2 OS) and U‐2 OS cells that express the three RPA subunits (SuperRPA). Graphic representation (related to Fig ) of the medians of CldU/IdU ratios in three biological replicates, the average is indicated in red.

Article Snippet: Immunoprecipitations were performed overnight at 4°C with protein G Dynabeads (Thermo Fisher Scientific) coupled to either rabbit immunoglobulin G (IgG) (P120‐201, Bethyl Laboratories), rabbit GNL3 (sc‐166460, Santa Cruz Biotechnology) or rabbit ORC2 antibody (A302‐734A, Bethyl Laboratories).

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

Journal: EMBO Reports

Article Title: The nucleolar protein GNL3 prevents resection of stalled replication forks

doi: 10.15252/embr.202357585

Figure Lengend Snippet: Immunofluorescence experiment in Flp‐In T‐Rex HEK293 cells showing the expression of GNL3‐BirA‐FLAG and the biotinylation by BirA (revealed by streptavidin coupled with Alexa‐488) upon addition of biotin. Common hits found by mass spectrometry between ORC2 immunoprecipitation and GNL3 BioID. The localization was determined using The Human Protein Atlas database ( https://www.proteinatlas.org/ ). Examples of GNL3 peaks on chromosome 19 obtained by ChIP‐seq of GNL3, INPUT is shown as negative control. The ORC2 ChIP‐seq data are obtained from (Miotto et al , ). PLA (proximity ligation assay) analyzing the proximity between ORC2 and GNL3 in HeLa S3 cells. PLA (proximity ligation assay) analyzing the proximity between GNL3 and CENP‐A in HeLa S3 cells.

Article Snippet: Immunoprecipitations were performed overnight at 4°C with protein G Dynabeads (Thermo Fisher Scientific) coupled to either rabbit immunoglobulin G (IgG) (P120‐201, Bethyl Laboratories), rabbit GNL3 (sc‐166460, Santa Cruz Biotechnology) or rabbit ORC2 antibody (A302‐734A, Bethyl Laboratories).

Techniques: Immunofluorescence, Expressing, Mass Spectrometry, Immunoprecipitation, ChIP-sequencing, Negative Control, Proximity Ligation Assay

Journal: EMBO Reports

Article Title: The nucleolar protein GNL3 prevents resection of stalled replication forks

doi: 10.15252/embr.202357585

Figure Lengend Snippet: GNL3‐BioID experiment analyzed by mass spectrometry. Expression of GNL3‐BirA‐FLAG in HEK293 Flp‐in cells was induced with doxycycline for 16 h then biotin was added for 4 h. For negative controls cells were treated 16 h with doxycycline alone or 4 h with biotin alone. Four biological replicates were analyzed by mass spectrometry. Label‐free quantification was performed using MaxQuant (Cox & Mann, ) and statistical analysis using Perseus (Tyanova et al , ). The volcano plot shows the proteins that are significantly (two‐tailed t ‐test, false discovery rate = 0.05) enriched upon induction of GNL3‐BirA‐FLAG and addition of biotin. The full list of proteins is available in Dataset . Western‐blot analysis of GNL3 and ORC2 immunoprecipitates in K562 cells. Comparison of the genomic location of GNL3 and ORC2. Chromatin immunoprecipitation of GNL3 followed by deep sequencing was performed in HeLa S3. GNL3‐binding sites were compared to ORC2‐binding sites obtained from Miotto et al . PLA (proximity ligation assay) analyzing the proximity between ORC2 and GNL3 in HeLa S3 cells that are stained with an antibody directed against NOP1. PLA (proximity ligation assay) analyzing the proximity between ORC2 and CENP‐A in HeLa S3 cells using the indicated antibodies. Graphic representation of the average number of PLA ORC2‐CENP‐A foci in three biological replicates. For statistical analysis, paired t ‐test was used; * P < 0.05. Source data are available online for this figure.

Article Snippet: Immunoprecipitations were performed overnight at 4°C with protein G Dynabeads (Thermo Fisher Scientific) coupled to either rabbit immunoglobulin G (IgG) (P120‐201, Bethyl Laboratories), rabbit GNL3 (sc‐166460, Santa Cruz Biotechnology) or rabbit ORC2 antibody (A302‐734A, Bethyl Laboratories).

Techniques: Mass Spectrometry, Expressing, Quantitative Proteomics, Two Tailed Test, Western Blot, Comparison, Chromatin Immunoprecipitation, Sequencing, Binding Assay, Proximity Ligation Assay, Staining

Journal: EMBO Reports

Article Title: The nucleolar protein GNL3 prevents resection of stalled replication forks

doi: 10.15252/embr.202357585

Figure Lengend Snippet: Schematic representation of human GNL3 protein with its associated domains (B: basic domain; C: coiled‐coil domain; G1: GTP‐binding motif 1; G4: GTP‐binding motif 4; I: intermediate domain; A: acidic domain). GNL3‐WT and GNL3‐dB are fused with FLAG. Western‐blot analysis of Flp‐In T‐Rex HeLa cells expressing exogenous GNL3‐WT or GNL3‐dB. Cells were transfected with siControl or siGNL3 for 48 h then expression of exogenous GNL3‐FLAG (resistant to the siRNA against GNL3) was induced using 10 μg/ml of doxycycline for 16 h. Immunofluorescence analysis of Flp‐in T‐Rex HeLa cells expressing exogenous GNL3‐WT or GNL3‐dB. PLA (proximity ligation assay) analyzing the proximity between ORC2 and GNL3‐FLAG or GNL3‐dB‐FLAG in HeLa Flp‐In cells upon doxycycline induction. Graphic representation of the average number of PLA ORC2‐FLAG foci in three biological replicates. For statistical analysis paired t ‐test was used; * P < 0.05. Immunofluorescence experiment of HeLa Flp‐In cells expressing GNL3‐dB with or without pre‐extraction with cytoskeletal buffer (CSK). Analysis of GIFD (Global Instant Fork Density) by DNA combing in HeLa cells ( n = 1; a biological replicate is shown in Fig ). GIFD value is indicated in red. Source data are available online for this figure.

Article Snippet: Immunoprecipitations were performed overnight at 4°C with protein G Dynabeads (Thermo Fisher Scientific) coupled to either rabbit immunoglobulin G (IgG) (P120‐201, Bethyl Laboratories), rabbit GNL3 (sc‐166460, Santa Cruz Biotechnology) or rabbit ORC2 antibody (A302‐734A, Bethyl Laboratories).

Techniques: Binding Assay, Western Blot, Expressing, Transfection, Immunofluorescence, Proximity Ligation Assay, Extraction

Journal: EMBO Reports

Article Title: The nucleolar protein GNL3 prevents resection of stalled replication forks

doi: 10.15252/embr.202357585

Figure Lengend Snippet: PLA (proximity ligation assay) analyzing the proximity between ORC2 and GNL3‐WT‐FLAG or GNL3‐dB‐FLAG in HeLa Flp‐In cells upon doxycycline induction using the indicated antibodies. PLA (proximity ligation assay) analyzing the proximity between ORC2 and GNL3‐dB‐FLAG in HeLa Flp‐In cells upon doxycycline induction using the indicated antibodies. Biological replicate of the GIFD (Global Instant Fork Density) analysis performed in Fig . GIFD value is indicated in red. Graphic representation (related to Fig ) of the medians of CldU/IdU ratios in three biological replicates, the average is indicated in red. Graphic representation (related to Fig ) of the medians of CldU/IdU ratios in three biological replicates, the average is indicated in red. Western‐blot analysis of Flp‐In T‐Rex HeLa cells expressing exogenous GNL3‐WT or GNL3‐dB mutants. Cells were transfected with siControl or siGNL3 for 48 h then the expression of GNL3‐WT and GNL3‐dB (resistant to the siRNA against GNL3) was induced using the indicated doses of doxycycline for 16 h. Immunofluorescence analysis of Flp‐in T‐Rex HeLa cells expressing GNL3‐dB at the indicated doses of doxycycline. Graphic representation (related to Fig ) of the medians of CldU/IdU ratios in three biological replicates, the average is indicated in red. Immunofluorescence experiment using the indicated antibodies of Flp‐in T‐Rex HeLa cells expressing or not exogenous FLAG‐ORC2. Graphic representation (related to Fig ) of the medians of CldU/IdU ratios in three biological replicates, the average is indicated in red.

Article Snippet: Immunoprecipitations were performed overnight at 4°C with protein G Dynabeads (Thermo Fisher Scientific) coupled to either rabbit immunoglobulin G (IgG) (P120‐201, Bethyl Laboratories), rabbit GNL3 (sc‐166460, Santa Cruz Biotechnology) or rabbit ORC2 antibody (A302‐734A, Bethyl Laboratories).

Techniques: Proximity Ligation Assay, Western Blot, Expressing, Transfection, Immunofluorescence

Journal: EMBO Reports

Article Title: The nucleolar protein GNL3 prevents resection of stalled replication forks

doi: 10.15252/embr.202357585

Figure Lengend Snippet: Flp‐in T‐Rex HeLa cells were sequentially labeled for 30 min with IdU and for 30 min with CldU then treated with 5 mM HU for 240 min. The ratio between CldU and IdU is plotted, the red line indicates the median. For statistical analysis Mann–Whitney test was used; **** P < 0.0001; *** P < 0.001; ns non‐significant. 100 individual DNA fibers were counted for each condition, biological replicates are shown in Fig . Flp‐in T‐Rex HeLa cells were sequentially labeled for 30 min with IdU and for 30 min with CldU then treated with 5 mM HU for 240 min with or without 10 μM of CDC7 inhibitor PHA‐767491. The ratio between CldU and IdU is plotted, the red line indicates the median. For statistical analysis Mann–Whitney test was used; **** P < 0.0001. 100 individual DNA fibers were counted for each condition, biological replicates are shown in Fig . Flp‐in T‐Rex HeLa cells were sequentially labeled for 30 min with IdU and for 30 min with CldU then treated with 5 mM HU for 240 min. The ratio between CldU and IdU is plotted, the red line indicates the median. For statistical analysis Mann–Whitney test was used; **** P < 0.0001; *** P < 0.001; ns non‐significant. 100 individual DNA fibers were counted for each condition, biological replicates are shown in Fig . PLA (proximity ligation assay) analyzing the proximity between ORC2 and GNL3‐dB‐FLAG in HeLa Flp‐In cells treated with indicated doses of doxycycline and stained with an antibody directed against NOP1. Flp‐in T‐Rex HeLa cells were sequentially labeled for 30 min with IdU and for 30 min with CldU then treated with 5 mM HU for 240 min. The ratio between CldU and IdU is plotted, the red line indicates the median. For statistical analysis, Mann–Whitney test was used; **** P < 0.0001. 100 individual DNA fibers were counted for each condition, and biological replicates are shown in Fig . Source data are available online for this figure.

Article Snippet: Immunoprecipitations were performed overnight at 4°C with protein G Dynabeads (Thermo Fisher Scientific) coupled to either rabbit immunoglobulin G (IgG) (P120‐201, Bethyl Laboratories), rabbit GNL3 (sc‐166460, Santa Cruz Biotechnology) or rabbit ORC2 antibody (A302‐734A, Bethyl Laboratories).

Techniques: Labeling, MANN-WHITNEY, Proximity Ligation Assay, Staining

Journal: EMBO Reports

Article Title: The nucleolar protein GNL3 prevents resection of stalled replication forks

doi: 10.15252/embr.202357585

Figure Lengend Snippet: HU‐treatment stalls active replication forks and activate dormant origins. When GNL3 is impaired, more origins may be licensed increasing the probability of replication origin firing. This could lead to over replication and replication stress. When GNL3‐deficient cells are treated with HU, the number of targets for HU is increased leading to activation of more dormant origins, a situation that leads to the exhaustion of the pool of available RPA that may explain the resection of nascent DNA. Inhibition of origin firing with CDC7 inhibitor, roscovitine or depletion of MCM3 counteracts this effect. Thanks to its ability to accumulate into the nucleolus, GNL3 may sequester a subset of ORC2 to precisely regulate origin licensing. When GNL3 is depleted (siGNL3) or does not accumulate in the nucleolus, ORC2 is released into the nucleoplasm leading to more licensed origins and DNA resection upon HU treatment.

Article Snippet: Immunoprecipitations were performed overnight at 4°C with protein G Dynabeads (Thermo Fisher Scientific) coupled to either rabbit immunoglobulin G (IgG) (P120‐201, Bethyl Laboratories), rabbit GNL3 (sc‐166460, Santa Cruz Biotechnology) or rabbit ORC2 antibody (A302‐734A, Bethyl Laboratories).

Techniques: Activation Assay, Inhibition

Journal: Cell reports

Article Title: TRF2 Mediates Replication Initiation within Human Telomeres to Prevent Telomere Dysfunction

doi: 10.1016/j.celrep.2020.108379

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-ORC2 , Bethyl , Cat# A302-735A: RRID:AB_10627808.

Techniques: Plasmid Preparation, Virus, Recombinant, Transfection, Blocking Assay, Purification, Gel Extraction, DNA Purification, shRNA, Software

Journal: Cell reports

Article Title: TRF2 Mediates Replication Initiation within Human Telomeres to Prevent Telomere Dysfunction

doi: 10.1016/j.celrep.2020.108379

Figure Lengend Snippet: (A) Doxycycline (Dox)-inducible expression of Myc-tagged TRF2ΔB. rtTA, reverse tetracycline trans -activator; TRE, tetracycline response element. (B) Immunoblot of cell lysates of LOX cell line stably expressing Dox-inducible TRF2ΔB. Untagged endogenous WT TRF2 (upper band) and Myc-tagged TRF2ΔB (lower band) are indicated. (C) ChIP analysis of LOX TRF2ΔB cells grown in the absence (−) or presence (+) of 1 μg/mL Dox for 4 days using antibodies against TRF2, ORC2, MCM3, H3K9me3, or immunoglobulin G (IgG) (control). Blots were probed by hybridization with 32 P-labeled telomeric TTAGGG (TelG) or Alu repeat (Alu) probes. Long Exp, long exposure. (D) Quantification of at least three independent experiments represented in (C). The ChIP data were first normalized to input, and the percent input for Dox (+) is shown as relative to dox (−), which was set as 1. Student’s t test was used for statistical analysis. Error bars indicate SD. **p < 0.01, ***p < 0.001; ns, no statistical significance (p > 0.05). (E) SMARD analysis of the Ch7q telomere segment from LOX TRF2ΔB cells grown for 3 days in the presence of 1 μg/mL Dox. Alignments of replicated molecules fully labeled with IdU (red) and CldU (green) are shown, collected from six independent samples stretched on slides (76 fully red- and 79 fully green-labeled molecules were also collected). Vertical lines (orange and blue) demarcate the boundaries where FISH probes bind, as described in . Symbols are as in . A replication profile histogram is shown under the molecule alignment.

Article Snippet: Antibodies used in ChIP assays include rabbit polyclonal antibodies ORC2 (Bethyl), MCM3 (Abcam), histone H3K9me3 (Diagenode) or IgG (Cell Signaling).

Techniques: Expressing, Western Blot, Stable Transfection, Control, Hybridization, Labeling

Journal: Cell reports

Article Title: TRF2 Mediates Replication Initiation within Human Telomeres to Prevent Telomere Dysfunction

doi: 10.1016/j.celrep.2020.108379

Figure Lengend Snippet: (A) Immunoblot of cell lysates of LOX cell lines stably expressing Dox-inducible SNF2H shRNA or Luc control shRNA. (B) ChIP analysis of LOX cells stably expressing Dox-inducible SNF2H shRNA grown in the absence+ (−) or presence (+) of 1 μg/mL Dox for 10 days using antibodies against ORC2, MCM3, or IgG (control). Blots were probed by hybridization with 32 P-labeled TelG or Alu probes. (C) Quantification of four independent experiments represented in (B). The ChIP data were first normalized to input, and the percent input for Dox (+) is shown as relative to Dox (−), which was set as 1. Student’s t test was used for statistical analysis. Error bars indicate SD. **p < 0.002; ns, p > 0.05. (D) Telomere length analysis of LOX cell lines stably expressing Dox-inducible SNF2H shRNA or Luc control shRNA. Cells were grown in the absence (−) or presence (+) of 1 μg/mL Dox for 10 days. Telomere length and relative amount of telomeric DNA were determined by restriction digestion of genomic DNA with AluI/MboI, followed by PFGE and Southern hybridization with a 32 P-labeled (CCCTAA) 4 probe. Ethidium bromide staining of the total DNA digest was used to normalize for DNA loading (bottom). Fragment size (in kilobases) is indicated on the left of the blot. The relative intensity of telomeric DNA signals (− Dox versus + Dox) is indicated below the blot. (E) Model of telomeric origin function in telomere replication. The repetitive sequence of telomeric DNA impedes and stalls replication forks. Failure to restart replication (i) leads to incomplete replication, telomere repeat loss, and dysfunction. Restart of replication from telomeric origins (ii) results in completion of telomere replication and proper telomere maintenance. (F) Model of telomeric origin assembly and firing. (i) SNF2H remodels telomere chromatin to allow ORC loading. (ii) TRF2 recruits the ORC. (iii) The ORC recruits cdc6 and cdt1, followed by double-hexamer MCM replicative helicase loading, resulting in pre-RC formation (origin licensing). (iv) Telomeric origin fires.

Article Snippet: Antibodies used in ChIP assays include rabbit polyclonal antibodies ORC2 (Bethyl), MCM3 (Abcam), histone H3K9me3 (Diagenode) or IgG (Cell Signaling).

Techniques: Western Blot, Stable Transfection, Expressing, shRNA, Control, Hybridization, Labeling, Staining, Sequencing

Journal: Cell reports

Article Title: TRF2 Mediates Replication Initiation within Human Telomeres to Prevent Telomere Dysfunction

doi: 10.1016/j.celrep.2020.108379

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Antibodies used in ChIP assays include rabbit polyclonal antibodies ORC2 (Bethyl), MCM3 (Abcam), histone H3K9me3 (Diagenode) or IgG (Cell Signaling).

Techniques: Plasmid Preparation, Virus, Recombinant, Transfection, Blocking Assay, Purification, Gel Extraction, DNA Purification, shRNA, Software