primary antibodies against mre11, brca1 and rpa70  (Novus Biologicals)

Journal: Cell Death & Disease

Article Title: HAT1 functions as a lactyltransferase and mediates RPA1 lactylation to promote DNA repair and radioresistance in lung adenocarcinoma

doi: 10.1038/s41419-025-08113-x

Figure Lengend Snippet: A The level of lactylated RPA1 in mutant cells with RPA1 K88R, K163R, K167R, and K267R. B The level of RPA1 interacting ssDNA in mutant cells with RPA1 K88R, K163R, K167R and K267R. C The level of RPA1 interacted with HR factors, including BLM, MRE11, RAD51, and NBS1, in mutant cells with RPA1 K88R, K163R, K167R, and K267R. D Relative HR efficiency in mutant cells with RPA1 K88R, K163R, K167R and K267R. All data were analyzed using two-way analysis of variance (ANOVA), ** P < 0.01.

Article Snippet: Antibodies against HAT1 (#11432-1-AP), β-actin (#66009-1-Ig), GAPDH (#60004-1-Ig), FLAG (#66008-4-Ig), HA (#51064-2-AP), BLM (#30254-1-AP), MRE11 (#10744-1-AP), NBS1 (#55025-1-AP) were obtained from Proteintech Group.

Techniques: Mutagenesis

Journal: Frontiers in Cell and Developmental Biology

Article Title: MRE11 orchestrates porcine oocyte meiotic progression by modulating the spindle assembly checkpoint

doi: 10.3389/fcell.2025.1635110

Figure Lengend Snippet: Subcellular localization and protein expression of Mre11 during porcine meiotic maturation. (A) Representative immunofluorescence images showing the localization of endogenous Mre11 in porcine oocytes at different developmental stages. Oocytes were immunostained with an anti-Mre11 antibody and counterstained with Hoechst. Scale bar, 2.5 μm. (B) Representative image showing the co-localization of Mre11 with kinetochores in porcine oocytes at the pro-metaphase I (Pro-M I) stage. Oocytes were immunostained with anti-Mre11 and CREST antibodies, and counterstained with Hoechst. Scale bar, 2.5 μm.

Article Snippet: Rabbit polyclonal anti-MRE11 antibody was purchased from Proteintech Group (Rosemont, IL, United States; Cat# 16370-1-AP); rabbit monoclonal anti-BUBR1 antibody was purchased from Abcam (Cambridge, United Kingdom, United States; Cat# ab133699); Mouse anti-α-tubulin-FITC antibody (Cat# F2168), mouse monoclonal anti-acetylation-α-tubulin antibody (T7451), peanut agglutinin (PNA)-FITC (L7381) and DAPI (D9542) were purchased from Sigma (Aldrich, St Louis, MO, United States); Rabbit polyclonal anti-γ-H2AX antibody (Cat# ab26350) was purchased from Abcam (Cambridge, United Kingdom); TRITC Phalloidin TRITC antibody (Cat# MX4405) was purchased from Maokang Biotechnology (Shanghai, China); Furthermore, goat anti-mouse IgG Alexa Fluor 488 antibody (Cat# A11029), donkey anti-Sheep IgG Alexa Fluor 594 antibody (Cat# A11016) were purchased from Thermo Fisher (Waltham, MA, United States). human anti-centromere antibody was purchased from Antibodies Incorporated (Davis, CA, United States; Cat# CA95617); rabbit monoclonal anti-GAPDH antibody was purchased from Cell Signaling Technology (Danvers, MA, United States; Cat# 2118).

Techniques: Expressing, Immunofluorescence

Journal: Frontiers in Cell and Developmental Biology

Article Title: MRE11 orchestrates porcine oocyte meiotic progression by modulating the spindle assembly checkpoint

doi: 10.3389/fcell.2025.1635110

Figure Lengend Snippet: Effect of Mre11 inhibition on porcine oocyte meiotic progression. (A) Representative images of cumulus cell expansion and polar body extrusion in control and Mre11-inhibited oocytes cultured in vitro for 44 h. Oocytes were denuded after culture to observe polar body extrusion. COCs: cumulus-oocyte complexes; DOs: denuded oocytes. Scale bars, 500 μm (a, e); 200 μm (b, f); 250 μm (c, g); 30 μm (d, h). (B) The rate of polar body extrusion was recorded in control and Mre11-inhibited groups treated with different concentrations (20 μM, 40 μM, 60 μM, and 100 μM) after 44 h of culture. (C) Representative images showing chromosome morphology at different developmental stages of oocyte maturation. DNA was counterstained with propidium iodide (PI). Scale bar, 5 μm. (D) The percentage of oocytes at different developmental stages was quantified in control and Mre11-inhibited groups. Data in panels (B) and (D) are presented as mean percentages (mean ± SEM) from at least three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Rabbit polyclonal anti-MRE11 antibody was purchased from Proteintech Group (Rosemont, IL, United States; Cat# 16370-1-AP); rabbit monoclonal anti-BUBR1 antibody was purchased from Abcam (Cambridge, United Kingdom, United States; Cat# ab133699); Mouse anti-α-tubulin-FITC antibody (Cat# F2168), mouse monoclonal anti-acetylation-α-tubulin antibody (T7451), peanut agglutinin (PNA)-FITC (L7381) and DAPI (D9542) were purchased from Sigma (Aldrich, St Louis, MO, United States); Rabbit polyclonal anti-γ-H2AX antibody (Cat# ab26350) was purchased from Abcam (Cambridge, United Kingdom); TRITC Phalloidin TRITC antibody (Cat# MX4405) was purchased from Maokang Biotechnology (Shanghai, China); Furthermore, goat anti-mouse IgG Alexa Fluor 488 antibody (Cat# A11029), donkey anti-Sheep IgG Alexa Fluor 594 antibody (Cat# A11016) were purchased from Thermo Fisher (Waltham, MA, United States). human anti-centromere antibody was purchased from Antibodies Incorporated (Davis, CA, United States; Cat# CA95617); rabbit monoclonal anti-GAPDH antibody was purchased from Cell Signaling Technology (Danvers, MA, United States; Cat# 2118).

Techniques: Inhibition, Control, Cell Culture, In Vitro

Journal: Frontiers in Cell and Developmental Biology

Article Title: MRE11 orchestrates porcine oocyte meiotic progression by modulating the spindle assembly checkpoint

doi: 10.3389/fcell.2025.1635110

Figure Lengend Snippet: Effect of Mre11 inhibition on BubR1 localization in porcine oocytes. (A) Representative images showing the localization of BubR1 at the metaphase I stage in control and Mre11-inhibited oocytes. Scale bar, 5 μm. (B) Quantitative analysis of BubR1 fluorescence intensity in control and Mre11-inhibited oocytes. Data are presented as the mean percentage (mean ± SEM) from at least three independent experiments. **P < 0.001.

Article Snippet: Rabbit polyclonal anti-MRE11 antibody was purchased from Proteintech Group (Rosemont, IL, United States; Cat# 16370-1-AP); rabbit monoclonal anti-BUBR1 antibody was purchased from Abcam (Cambridge, United Kingdom, United States; Cat# ab133699); Mouse anti-α-tubulin-FITC antibody (Cat# F2168), mouse monoclonal anti-acetylation-α-tubulin antibody (T7451), peanut agglutinin (PNA)-FITC (L7381) and DAPI (D9542) were purchased from Sigma (Aldrich, St Louis, MO, United States); Rabbit polyclonal anti-γ-H2AX antibody (Cat# ab26350) was purchased from Abcam (Cambridge, United Kingdom); TRITC Phalloidin TRITC antibody (Cat# MX4405) was purchased from Maokang Biotechnology (Shanghai, China); Furthermore, goat anti-mouse IgG Alexa Fluor 488 antibody (Cat# A11029), donkey anti-Sheep IgG Alexa Fluor 594 antibody (Cat# A11016) were purchased from Thermo Fisher (Waltham, MA, United States). human anti-centromere antibody was purchased from Antibodies Incorporated (Davis, CA, United States; Cat# CA95617); rabbit monoclonal anti-GAPDH antibody was purchased from Cell Signaling Technology (Danvers, MA, United States; Cat# 2118).

Techniques: Inhibition, Control, Fluorescence

Journal: Frontiers in Cell and Developmental Biology

Article Title: MRE11 orchestrates porcine oocyte meiotic progression by modulating the spindle assembly checkpoint

doi: 10.3389/fcell.2025.1635110

Figure Lengend Snippet: Effect of Mre11 inhibition on spindle assembly and chromosome alignment in porcine oocytes. (A) Representative images depicting spindle morphologies and chromosome alignment in control and Mre11-inhibited oocytes. Metaphase I (M I) oocytes were immunostained with anti-α-tubulin-FITC antibody to visualize spindles and counterstained with propidium iodide (PI) to visualize chromosomes. Scale bar, 5 μm. (B) The frequency of aberrant spindles was recorded in control and Mre11-inhibited oocytes. Data in panel (B) are presented as mean percentages (mean ± SEM) from at least three independent experiments.

Article Snippet: Rabbit polyclonal anti-MRE11 antibody was purchased from Proteintech Group (Rosemont, IL, United States; Cat# 16370-1-AP); rabbit monoclonal anti-BUBR1 antibody was purchased from Abcam (Cambridge, United Kingdom, United States; Cat# ab133699); Mouse anti-α-tubulin-FITC antibody (Cat# F2168), mouse monoclonal anti-acetylation-α-tubulin antibody (T7451), peanut agglutinin (PNA)-FITC (L7381) and DAPI (D9542) were purchased from Sigma (Aldrich, St Louis, MO, United States); Rabbit polyclonal anti-γ-H2AX antibody (Cat# ab26350) was purchased from Abcam (Cambridge, United Kingdom); TRITC Phalloidin TRITC antibody (Cat# MX4405) was purchased from Maokang Biotechnology (Shanghai, China); Furthermore, goat anti-mouse IgG Alexa Fluor 488 antibody (Cat# A11029), donkey anti-Sheep IgG Alexa Fluor 594 antibody (Cat# A11016) were purchased from Thermo Fisher (Waltham, MA, United States). human anti-centromere antibody was purchased from Antibodies Incorporated (Davis, CA, United States; Cat# CA95617); rabbit monoclonal anti-GAPDH antibody was purchased from Cell Signaling Technology (Danvers, MA, United States; Cat# 2118).

Techniques: Inhibition, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: MRE11 orchestrates porcine oocyte meiotic progression by modulating the spindle assembly checkpoint

doi: 10.3389/fcell.2025.1635110

Figure Lengend Snippet: Effect of Mre11 inhibition on the acetylation level of α-tubulin in porcine oocytes. (A) Representative images of microtubule fibers before and after nocodazole treatment in control and Mre11-inhibited oocytes. Oocytes were immunostained with an anti-α-tubulin-FITC antibody to visualize microtubules and counterstained with propidium iodide (PI) to visualize chromosomes. Scale bar, 10 μm. (B) Representative images showing the acetylation of α-tubulin in control and Mre11-inhibited oocytes. Oocytes were immunostained with an anti-acetyl-α-tubulin (Lys-40) antibody to assess the acetylation level of α-tubulin. Scale bar, 5 μm. (C) The fluorescence intensity of acetylated α-tubulin was quantified in control and Mre11-inhibited oocytes. Data in panel (C) are presented as mean percentages (mean ± SEM) from at least three independent experiments. ***P < 0.001.

Article Snippet: Rabbit polyclonal anti-MRE11 antibody was purchased from Proteintech Group (Rosemont, IL, United States; Cat# 16370-1-AP); rabbit monoclonal anti-BUBR1 antibody was purchased from Abcam (Cambridge, United Kingdom, United States; Cat# ab133699); Mouse anti-α-tubulin-FITC antibody (Cat# F2168), mouse monoclonal anti-acetylation-α-tubulin antibody (T7451), peanut agglutinin (PNA)-FITC (L7381) and DAPI (D9542) were purchased from Sigma (Aldrich, St Louis, MO, United States); Rabbit polyclonal anti-γ-H2AX antibody (Cat# ab26350) was purchased from Abcam (Cambridge, United Kingdom); TRITC Phalloidin TRITC antibody (Cat# MX4405) was purchased from Maokang Biotechnology (Shanghai, China); Furthermore, goat anti-mouse IgG Alexa Fluor 488 antibody (Cat# A11029), donkey anti-Sheep IgG Alexa Fluor 594 antibody (Cat# A11016) were purchased from Thermo Fisher (Waltham, MA, United States). human anti-centromere antibody was purchased from Antibodies Incorporated (Davis, CA, United States; Cat# CA95617); rabbit monoclonal anti-GAPDH antibody was purchased from Cell Signaling Technology (Danvers, MA, United States; Cat# 2118).

Techniques: Inhibition, Control, Fluorescence

Journal: Frontiers in Cell and Developmental Biology

Article Title: MRE11 orchestrates porcine oocyte meiotic progression by modulating the spindle assembly checkpoint

doi: 10.3389/fcell.2025.1635110

Figure Lengend Snippet: Effect of Mre11 inhibition on cytoplasmic actin assembly in porcine oocytes. (A) Representative images showing the distribution of actin filaments at the oocyte cortex and cytoplasm in the control and Mre11-inhibited groups. Red indicates F-actin. Scale bar, 20 μm. (B) Quantification of fluorescence intensity of F-actin at the cortex in the control and Mre11-inhibited groups. (C) Quantification of fluorescence intensity of F-actin in the cytoplasm in the control and Mre11-inhibited groups. (D) Quantitative analysis of F-actin fluorescence intensities at the cortex and in the cytoplasm. Data in panels (B–D) are presented as mean percentages (mean ± SEM) from at least three independent experiments. ***P < 0.001.

Article Snippet: Rabbit polyclonal anti-MRE11 antibody was purchased from Proteintech Group (Rosemont, IL, United States; Cat# 16370-1-AP); rabbit monoclonal anti-BUBR1 antibody was purchased from Abcam (Cambridge, United Kingdom, United States; Cat# ab133699); Mouse anti-α-tubulin-FITC antibody (Cat# F2168), mouse monoclonal anti-acetylation-α-tubulin antibody (T7451), peanut agglutinin (PNA)-FITC (L7381) and DAPI (D9542) were purchased from Sigma (Aldrich, St Louis, MO, United States); Rabbit polyclonal anti-γ-H2AX antibody (Cat# ab26350) was purchased from Abcam (Cambridge, United Kingdom); TRITC Phalloidin TRITC antibody (Cat# MX4405) was purchased from Maokang Biotechnology (Shanghai, China); Furthermore, goat anti-mouse IgG Alexa Fluor 488 antibody (Cat# A11029), donkey anti-Sheep IgG Alexa Fluor 594 antibody (Cat# A11016) were purchased from Thermo Fisher (Waltham, MA, United States). human anti-centromere antibody was purchased from Antibodies Incorporated (Davis, CA, United States; Cat# CA95617); rabbit monoclonal anti-GAPDH antibody was purchased from Cell Signaling Technology (Danvers, MA, United States; Cat# 2118).

Techniques: Inhibition, Control, Fluorescence

Journal: Frontiers in Cell and Developmental Biology

Article Title: MRE11 orchestrates porcine oocyte meiotic progression by modulating the spindle assembly checkpoint

doi: 10.3389/fcell.2025.1635110

Figure Lengend Snippet: Effect of Mre11 inhibition on meiotic spindle positioning and the formation of actin caps in porcine oocytes. (A) Representative images showing spindle positioning in control and Mre11-inhibited oocytes. (B) To quantify spindle position after Mre11 inhibition, D is defined as the distance from the proximal end of the spindle pole to the cortex, while L is defined as the distance from the distal end of the spindle pole to the cortex. (C) The D/L ratio in the Mre11-inhibited group was significantly higher than that in the control group. (D) Formation of actin caps and CGFDs. After Mre11 inhibition, the actin cap over a chromosome was disrupted, and Mre11 inhibition also affected CGFD formation. Red indicates actin; green indicates CGs; blue indicates chromatin. Scale bar, 20 μm.

Article Snippet: Rabbit polyclonal anti-MRE11 antibody was purchased from Proteintech Group (Rosemont, IL, United States; Cat# 16370-1-AP); rabbit monoclonal anti-BUBR1 antibody was purchased from Abcam (Cambridge, United Kingdom, United States; Cat# ab133699); Mouse anti-α-tubulin-FITC antibody (Cat# F2168), mouse monoclonal anti-acetylation-α-tubulin antibody (T7451), peanut agglutinin (PNA)-FITC (L7381) and DAPI (D9542) were purchased from Sigma (Aldrich, St Louis, MO, United States); Rabbit polyclonal anti-γ-H2AX antibody (Cat# ab26350) was purchased from Abcam (Cambridge, United Kingdom); TRITC Phalloidin TRITC antibody (Cat# MX4405) was purchased from Maokang Biotechnology (Shanghai, China); Furthermore, goat anti-mouse IgG Alexa Fluor 488 antibody (Cat# A11029), donkey anti-Sheep IgG Alexa Fluor 594 antibody (Cat# A11016) were purchased from Thermo Fisher (Waltham, MA, United States). human anti-centromere antibody was purchased from Antibodies Incorporated (Davis, CA, United States; Cat# CA95617); rabbit monoclonal anti-GAPDH antibody was purchased from Cell Signaling Technology (Danvers, MA, United States; Cat# 2118).

Techniques: Inhibition, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: MRE11 orchestrates porcine oocyte meiotic progression by modulating the spindle assembly checkpoint

doi: 10.3389/fcell.2025.1635110

Figure Lengend Snippet: Effect of Mre11 inhibition on DNA damage levels in porcine oocytes. (A) Representative images showing DNA damage in control and Mre11-inhibited oocytes. Oocytes were immunostained with an anti-γH2A.X antibody and counterstained with Hoechst. Scale bar, 5 μm. (B) Quantification of γH2A.X fluorescence intensity in control and Mre11-inhibited oocytes. Data are presented as mean percentages (mean ± SEM) from at least three independent experiments. ***P < 0.001.

Article Snippet: Rabbit polyclonal anti-MRE11 antibody was purchased from Proteintech Group (Rosemont, IL, United States; Cat# 16370-1-AP); rabbit monoclonal anti-BUBR1 antibody was purchased from Abcam (Cambridge, United Kingdom, United States; Cat# ab133699); Mouse anti-α-tubulin-FITC antibody (Cat# F2168), mouse monoclonal anti-acetylation-α-tubulin antibody (T7451), peanut agglutinin (PNA)-FITC (L7381) and DAPI (D9542) were purchased from Sigma (Aldrich, St Louis, MO, United States); Rabbit polyclonal anti-γ-H2AX antibody (Cat# ab26350) was purchased from Abcam (Cambridge, United Kingdom); TRITC Phalloidin TRITC antibody (Cat# MX4405) was purchased from Maokang Biotechnology (Shanghai, China); Furthermore, goat anti-mouse IgG Alexa Fluor 488 antibody (Cat# A11029), donkey anti-Sheep IgG Alexa Fluor 594 antibody (Cat# A11016) were purchased from Thermo Fisher (Waltham, MA, United States). human anti-centromere antibody was purchased from Antibodies Incorporated (Davis, CA, United States; Cat# CA95617); rabbit monoclonal anti-GAPDH antibody was purchased from Cell Signaling Technology (Danvers, MA, United States; Cat# 2118).

Techniques: Inhibition, Control, Fluorescence

Journal: NAR Cancer

Article Title: SRSF2 overexpression induces transcription-/replication-dependent DNA double-strand breaks and interferes with DNA repair pathways to promote lung tumor progression

doi: 10.1093/narcan/zcaf011

Figure Lengend Snippet: SRSF2 interacts with and increases MRE11 recruitment to chromatin. (A, B) Representative immunoblots of indicated proteins in WCEs (A, left panel) or chromatin-enriched fractions (B) from H358-SRSF2 cells cultured in the presence (+) or absence (−) of doxycycline (1 μg/ml) for 48 h. ( A ) Right panel: Relative quantification of MRE11/RAD50/NBS1 protein level normalized to α-tubulin. Ratio in uninduced condition was arbitrarily assigned the value 1. Mean ± SD. n = 3. Paired t -test, * P < .05, ns: not significant. ( B ) A representative experiment out of three is illustrated. Numbers represent the quantification of densitometric signals according to histone H3 signal. ( C ) Representative immunoblots of indicated proteins in A549 or H1299 cells transfected for 72 h either with control (Ctrl) or Srsf2 siRNA in chromatin-enriched fractions (upper panels) and WCEs. Numbers represent the quantification of the densitometric signals according to histone H3 signal. n = 3. ( D ) Co-IP of SRSF2-HA or SRSF2(P95H)-HA with MRE11, RAD50, or NBS1 proteins in A549 cells transiently transfected for 24 h with pcDNA3.1 (Ctrl), pcDN3.1-SRSF2-HA, or pcDNA3.1-SRSF2(P95H)-HA plasmid. Input represents 10% of the immunoprecipitates. A representative experiment out of three is presented. ( E ) Biolayer interferometry using recombinant MRE11 as a bait and in vitro transcribed/translated pcDNA3.1 (lysate), SRSF2-HA (SRSF2), or SRSF2(P95H)-HA (SRSF2-P95H) plasmid. Upper panel: Western blotting of in vitro transcribed/translated recombinant proteins using anti-HA antibody. Ctrl: pcDNA3.1. ( F ) Upper panel: PLA for the detection of endogenous SRSF2 and MRE11 interaction in A549 cells. Ctrl – (negative control): PLA with mouse anti-SRSF2 antibody only. Scale bar = 10 μm. Lower panel: Quantification (mean ± SD) of the number of PLA spots/nucleus. n = 3 performed in duplicate (30–40 cells/condition). Mann–Whitney t -test, ** P < .01. ( G ) Representative immunoblots of indicated proteins in chromatin-enriched or RIPA extracts from A549 cells transiently transfected for 24 h with the indicated constructs. n = 2. ( H ) Upper panels: PLA for the detection of endogenous SRSF2 and MRE11 interaction in A549 cells treated or not (NT) with DRB (50 μM) for 6 h. Ctrl – (negative control): PLA with mouse anti-SRSF2 antibody only. SRSF2/MRE11: PLA with mouse anti-SRSF2 and rabbit anti-MRE11 antibodies. Scale bar = 10 μm. Lower panel: Quantification (mean ± SD) of the number of PLA spots/nucleus. n = 3 performed in duplicate (30–40 cells/condition). Mann–Whitney t -test, ** P < .01. In data, α-tubulin, and histone H3 were used as loading controls for whole cell and chromatin-enriched fractions, respectively.

Article Snippet: Antibody for MRE11 immunostaining was mouse monoclonal anti-MRE11 (clone 12D7, cat. #GTX70212, Euromedex, Souffleweyersheim, France).

Techniques: Western Blot, Cell Culture, Quantitative Proteomics, Transfection, Control, Co-Immunoprecipitation Assay, Plasmid Preparation, Recombinant, In Vitro, Negative Control, MANN-WHITNEY, Construct

Journal: NAR Cancer

Article Title: SRSF2 overexpression induces transcription-/replication-dependent DNA double-strand breaks and interferes with DNA repair pathways to promote lung tumor progression

doi: 10.1093/narcan/zcaf011

Figure Lengend Snippet: SRSF2 promotes DNA repair by HR. ( A ) H358-SRSF2 cells were transfected with mismatch (ctrl ) , mre11 , nbs1 , or rad50 siRNA for 48 h and treated (+) or not (−) with doxycycline (Dox) (1 μg/ml) for additional 24 h. Representative immunoblots of indicated proteins are shown. Numbers represent densitometric quantification of specific protein signal normalized to α-tubulin signal. n = 3. (B–D) Quantification of DSB-induced HR in H1299 clones stably expressing the pDR-GFP (pBL174) plasmid. When these cells are transfected with the pBL133 plasmid encoding the I-SceI restriction enzyme, efficient recombination restores a functional GFP-coding sequence. ( B ) Quantification of DSB-induced HR in H1299-pBL174 cells transfected with control (Ctrl) or Srsf2 siRNAs and either pcDNA3.1 (−I-SceI) or pBL133 (+I-SceI). Upper panels: Representative FL1-H versus FSC-H dot plots of GFP-positive (FL1-H +) and –negative (FL1-H −) cells. Lower left panel: Western blotting of SRSF2 and I-SceI-HA. Lower right panel: Percentage of GFP-positive cells. n = 4. Unpaired t -test, *** P < .001, ** P < .01. ( C ) Quantification of DSB-induced HR in H1299-pBL174 cells co-transfected with pBL133 (+I-SceI) and either pcDNA3.1 (Ctrl) or pcDNA3.1-SRSF2-HA. Left panel: Western blotting of HA-tagged SRSF2 and I-SceI proteins. Right panel: Percentage of HA-/GFP-positive transfected cells determined by flow cytometry. n = 6. Paired t -test, * P < .05. ( D ) Quantification of DSB-induced HR in H1299-pBL174 cells co-transfected with pBL133 (+I-SceI) together with pcDNA3.1-SRSF2-HA (SRSF2) and treated or not (NT) with mirin (40 μM) or DRB (50 μM). Percentage of HA-/GFP-positive transfected cells was determined by flow cytometry. n = 4. Paired t -test, * P < .05. ( E ) H358-SRSF2 clones were cultured with (+) or without (−) doxycycline (Dox) (1 μg/ml) for 24 h. Upper panels: Representative images of RAD51 immunostaining. DAPI was used to counterstain nuclei. Scale bar = 10 μm. Lower panel: Quantification of the number of RAD51 foci/nucleus. Mean ± SD. n = 3. Mann–Whitney t -test, **** P < .0001. ( F , G ) H358-SRSF2 cells were cultured with (+) or without (−) doxycycline (Dox) (1 μg/ml) for 48 h in the presence or absence of mirin (40 μM, 24 h co-treatment, panel F) or DRB (50 μM, 6 h co-treatment, panel G). Left panels: Representative images of RPA IF. Scale bar = 20 μm. Right panels: Quantification (mean ± SD) of RPA-positive cells (%) in each condition. n = 3. Unpaired t -test, **** P < .0001, *** P <. 001, * *P < .01, * P < .05, ns: not significant. All data, α-Tubulin was used as a loading control.

Article Snippet: Antibody for MRE11 immunostaining was mouse monoclonal anti-MRE11 (clone 12D7, cat. #GTX70212, Euromedex, Souffleweyersheim, France).

Techniques: Transfection, Western Blot, Clone Assay, Stable Transfection, Expressing, Plasmid Preparation, Functional Assay, Sequencing, Control, Flow Cytometry, Cell Culture, Immunostaining, MANN-WHITNEY

Journal: NAR Cancer

Article Title: SRSF2 overexpression induces transcription-/replication-dependent DNA double-strand breaks and interferes with DNA repair pathways to promote lung tumor progression

doi: 10.1093/narcan/zcaf011

Figure Lengend Snippet: High level of both Srsf2 and Mre11 mRNAs correlates with poor prognosis in stage I LUAD patients. ( A ) Left panels: Representative MRE11 immunostainings from paraffin-embedded sections of LUAD patients. Immunoscores are indicated for each case. Right panel: Distribution of IHC scores of MRE11 and SRSF2 in NSCLC patients. ( B ) Spearman correlation between SRSF2 and MRE11 immunoscores. ( C ) Normalized Srsf2 (upper panel) and Mre11 (lower panel) mRNA levels [log2(FPKM-UQ + 1)] in LUAD patients and matched normal lung tissues retrieved from the TCGA ( n = 524 for primary LUAD tumors and n = 59 for matched normal tissues) using UCSC Xena platform. P -values were calculated using an unpaired t -test. ( D ) Relationship between normalized Srsf2 and Mre11 (upper panel) or 53BP1 (lower panel) mRNA levels in normal lung tissues and LUAD patients. Spearman correlation. ( E ) Kaplan–Meier univariate survival analysis of early stage (pTNM I) LUAD patients from TCGA displaying high level of Srsf2 mRNA (up to median, n = 134) and either high (>75th percentile, 4th quartile, n = 45) or low/medium (<75th percentile, 1st–3rd quartiles, n = 89) Mre11 mRNA levels. Log-rank test.

Article Snippet: Antibody for MRE11 immunostaining was mouse monoclonal anti-MRE11 (clone 12D7, cat. #GTX70212, Euromedex, Souffleweyersheim, France).

Techniques:

Journal: NAR Cancer

Article Title: SRSF2 overexpression induces transcription-/replication-dependent DNA double-strand breaks and interferes with DNA repair pathways to promote lung tumor progression

doi: 10.1093/narcan/zcaf011

Figure Lengend Snippet: Model for SRSF2-induced lung tumor progression. High level of SRSF2 induces a global transcriptional increase in LUAD cells. This leads to enhanced replicative stress and DNA DSBs, likely as a result of conflicts between transcription and replication machineries. SRSF2 also controls DSB repair. SRSF2 inhibits c-NHEJ that correlates with a decrease of 53BP1 mRNA and protein levels. Conversely, a cross talk between SRSF2 and MRE11 proteins, which are both commonly increased in LUAD patients and are found in closed proximity in LUAD cells, increases DSB repair by HR. We propose that a high level of SRSF2, along with MRE11, tightly controls the balance between DSB production and repair helping to maintain genome instability below a threshold compatible with tumor cell survival, and thus, contributing to lung tumor progression.

Article Snippet: Antibody for MRE11 immunostaining was mouse monoclonal anti-MRE11 (clone 12D7, cat. #GTX70212, Euromedex, Souffleweyersheim, France).

Techniques: