Journal: Cell reports

Article Title: BRCA1 Mutation-Specific Responses to 53BP1 Loss-Induced Homologous Recombination and PARP Inhibitor Resistance

doi: 10.1016/j.celrep.2018.08.086

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit Anti-BRCA2 Polyclonal antibody , Bethyl , Cat# A303–434A; RRID: AB_10952240.

Techniques: Labeling, Polymer, Recombinant, Plasmid Preparation, Staining, Mutagenesis, Extraction, Expressing, TaqMan Assay, shRNA, Software

Journal: Molecular medicine reports

Article Title: Effect of S100A8 and S100A9 on expressions of cytokine and skin barrier protein in human keratinocytes.

doi: 10.3892/mmr.2019.10454

Figure Lengend Snippet: Figure 6. HaCaT cells were treated with (A) S100A8 or (B) S100A9 (10 µg/ml) for 24 h and TLR4 expression was increased as assessed by western blotting. Data are expressed as the mean ± standard deviation. **P<0.01 vs. the control. S100, calcium binding protein; TLR4, toll‑like receptor 4.

Article Snippet: Tlr4 inhibitor cli-095 (Tlr4i), protein kinase δ (PKcδ) inhibitor (rottlerin), p38 mitogen-activated Correspondence to: dr in Sik Kim, department of Biomedical laboratory Science, School of Medicine, eulji university, 77 Gyeryoung-ro 771 beon-gil, Jung-Gu, daejeon 34824, republic of Korea e-mail: orientree@eulji.ac.kr Key words: atopic dermatitis, S100 calcium binding protein a8, S100 calcium binding protein a9, keratinocyte, skin barrier protein protein kinase (MaPK) inhibitor (SB202190), MeK inhibitor (Pd98059) and nuclear factor-κB (nF-κB) inhibitor (BaY-11-7085) were obtained from calbiochem (Merck KGaa). antibodies against p38 MaPK (cat. no. 9212), phospho-p38 MaPK (cat. no. 9211), phospho-extracellular signal regulated kinase 1/2 (erK1/2; cat. no. 9101), rabbit igG-HrP (cat. no. 7074), and mouse igG-HrP (cat. no. 7076) were acquired from cell Signaling Technology, inc. anti-erK2 (cat. no. sc-154) and anti-Tlr4 (cat. no. sc-10741) antibodies were obtained from Santa cruz Biotechnology, inc. Production of recombinant S100A8 and S100A9 proteins. in our previous report, the cdna of human S100a8 and S100a9 was cloned into peT28 expression vector (Merck KGaa) (10). recombinant S100a8 and S100a9 expression was induced with 1 mM isopropyl β-d-thiogalactoside in E. coli Bl21 (de3; Merck KGaa).

Techniques: Expressing, Western Blot, Standard Deviation, Control, Binding Assay

Journal: EMBO Reports

Article Title: Arginine methylation-dependent METTL14-SMN interaction regulates RNA m 6 A homeostasis

doi: 10.1038/s44319-025-00590-7

Figure Lengend Snippet: ( A ) Sanger sequencing confirmation of Mettl14 arginine methylation deficient mouse model. In total, 13 arginine resides located at the C-terminal disordered region of Mettl14 were mutated to lysine residues. ( B ) The arginine methylation levels of Mettl14 in WT, heterozygous (WT/RK), and homozygous (RK/RK) mice were detected by IP-western blot using tissue lysates from mouse spleen and thymus. ( C ) The protein expression levels of several DNA repair genes, including BRCA1, ATRIP, and PALB2, were detected by western blot using Thymus tissues from WT, WT/RK, and RK/RK mice. ( D ) Mettl14 arginine methylation deficiency increases the population of primitive hematopoietic cells (LK cells). ** P = 0.0019. ( E ) Mettl14 arginine methylation deficiency increases the population of myeloid (Mac1 + Gr1 + ) cells in the BM, but not in the spleen. * P = 0.013. ( F ) Mettl14 arginine methylation deficiency decreases the B lymphoid (B220 + ) population in the BM, but not in the spleen. * P = 0.011. ( G ) Mettl14 arginine methylation deficiency decreases the T lymphoid (CD3 + ) population in the BM, but not in the spleen. Data from three independent replicates were analyzed by Student’s t test and shown as mean ± SD. ** P = 0.0097. .

Article Snippet: Rabbit anti-PALB2 antibody , Proteintech , 14340-1-AP.

Techniques: Sequencing, Methylation, Western Blot, Expressing

Journal: Cell Death & Disease

Article Title: BRCA2 deficiency is a potential driver for human primary ovarian insufficiency

doi: 10.1038/s41419-019-1720-0

Figure Lengend Snippet: a The gene panel on the RT 2 Profiler PCR Array. These genes are crucial factors in BER, NER, MMR, DSB repair, or other relative pathways in DNA damage response. b Quality control test of the whole panel of the array. Five control genes (ACTB, B2M, GAPDH, HPRT1, and RPLPO) were contained on the array as quality control. Average CT value of the control genes were summarized from real-time qPCR of the array. All the included oocytes showed qualified CT value except oocyte POI-10 and oocyte POI-13. These two oocytes were thus excluded from the subsequent experiments. c Expression level of the 84 DNA damage response genes among the individual human oocytes. POI-04 and POI-20 oocytes showed significantly lower expression of BRCA2 , while POI-17 oocyte expressed lower XRCC2

Article Snippet: Rabbit polyclonal anti-Brca2 antibody was purchased from Santa Cruz Biotechnology (Dallas, TA, USA; Cat#: sc-293185); mouse monoclonal anti-α-tubulin-fluorescein isothiocyanate (FITC) antibody was purchased from Sigma (St. Louis, MO, USA; Cat#: F2168); Rabbit monoclonal anti-γH2AX and anti-Gapdh antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA; Cat#: 9718, 2118); HRP-conjugated secondary antibodies were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA, USA); FITC-conjugated goat anti-rabbit IgG (H + L), TRITC-conjugated goat anti-rabbit IgG (H + L) and FITC-conjugated goat anti-mouse IgG (H + L) were purchased from Zhongshan Golden Bridge Biotechnology Co., LTD (Beijing, China).

Techniques: Control, Expressing

Journal: Cell Death & Disease

Article Title: BRCA2 deficiency is a potential driver for human primary ovarian insufficiency

doi: 10.1038/s41419-019-1720-0

Figure Lengend Snippet: a Schematic illustration of deletion of Brca2 exons and creation of Brca2 knockout allele by Gdf9-Cre -mediated recombination in oocytes. b Immunoblotting analysis of Brca2 protein expression in control and Brca2 -deficient oocytes. The amount of Gapdh was used as an internal control. Molecular mass is given in kilodaltons. Two hundered germinal vesicle oocytes isolated from ovaries of 2-month-old mice were used for each lane of the blots. For each experiment, at least five mice of each genotype were used. c Comparison of breeding between control and Brca2 -deficient females. The female mice were cocaged with fertile male mice. The breeding assay was recorded for at least 6 months

Article Snippet: Rabbit polyclonal anti-Brca2 antibody was purchased from Santa Cruz Biotechnology (Dallas, TA, USA; Cat#: sc-293185); mouse monoclonal anti-α-tubulin-fluorescein isothiocyanate (FITC) antibody was purchased from Sigma (St. Louis, MO, USA; Cat#: F2168); Rabbit monoclonal anti-γH2AX and anti-Gapdh antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA; Cat#: 9718, 2118); HRP-conjugated secondary antibodies were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA, USA); FITC-conjugated goat anti-rabbit IgG (H + L), TRITC-conjugated goat anti-rabbit IgG (H + L) and FITC-conjugated goat anti-mouse IgG (H + L) were purchased from Zhongshan Golden Bridge Biotechnology Co., LTD (Beijing, China).

Techniques: Knock-Out, Western Blot, Expressing, Control, Isolation, Comparison

Journal: Cell Death & Disease

Article Title: BRCA2 deficiency is a potential driver for human primary ovarian insufficiency

doi: 10.1038/s41419-019-1720-0

Figure Lengend Snippet: a Representative images of ovarian size from control and Brca2 -deficient mice. Scale bar, 200 μm. b Average weights of ovaries obtained from control and Brca2 -deficient females. Data are shown as mean ± SEM. c – f Histology of ovarian sections from young adult control and Brca2 -deficient females. Ovaries from 2-month-old mice were embedded in paraffin, and sections of 8 μm thickness were prepared and stained with H&E. Black arrows in ( d ) show the growing follicles at different developmental stages; yellow arrows in ( f ) indicate the developmentally arrested follicles with degenerating oocytes. CL, corpus luteum. Scale bars in ( c ) and ( e ), 250 μm; scale bars in ( d ) and ( f ), 50 μm. g Quantification analysis of primordial follicles between control and Brca2 -deficient ovaries. h Quantification analysis of activated follicles between control and Brca2 -deficient ovaries. Brca2 -deficient ovary contained a significantly increased number of activated follicles with developmental arrest and degraded oocytes compared with the control ovary

Article Snippet: Rabbit polyclonal anti-Brca2 antibody was purchased from Santa Cruz Biotechnology (Dallas, TA, USA; Cat#: sc-293185); mouse monoclonal anti-α-tubulin-fluorescein isothiocyanate (FITC) antibody was purchased from Sigma (St. Louis, MO, USA; Cat#: F2168); Rabbit monoclonal anti-γH2AX and anti-Gapdh antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA; Cat#: 9718, 2118); HRP-conjugated secondary antibodies were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA, USA); FITC-conjugated goat anti-rabbit IgG (H + L), TRITC-conjugated goat anti-rabbit IgG (H + L) and FITC-conjugated goat anti-mouse IgG (H + L) were purchased from Zhongshan Golden Bridge Biotechnology Co., LTD (Beijing, China).

Techniques: Control, Staining

Journal: Cell Death & Disease

Article Title: BRCA2 deficiency is a potential driver for human primary ovarian insufficiency

doi: 10.1038/s41419-019-1720-0

Figure Lengend Snippet: a Representative images of in vivo matured eggs from control and Brca2 -deficient mice. Scale bar, 20μm. b The number of eggs obtained from control and Brca2 -deficient mice. c The rate of fragmented eggs retrieved from control and Brca2 -deficient mice. d Immunofluorescence of phosphorylated H2AX (γH2AX) in control and Brca2 -deficient oocytes. e Fluorescence intensity of γH2AX signal in control and Brca2 -deficient oocytes were summarized. f Representative images of GV oocytes obtained from control and Brca2 -deficient ovaries. Scale bar, 20 μm. g The rate of developmentally arrested oocytes in control and Brca2 -deficient mice. h Representative images of MII oocytes matured in vitro from control and Brca2 -deficient GV oocytes. Scale bar, 20 μm. i The rate of first polar body extrusion of control and Brca2 -deficient oocytes matured in vitro. j Representative images of spindle morphology and chromosome alignment in control and Brca2 -deficient oocytes after in vitro maturation. Oocytes were immunostained with anti-α-tubulin-FITC antibody to visualize spindles and counterstained with Hoechst to visualize chromosomes. Scale bar, 20 μm. k The percentage of oocytes with aberrant spindle in control and Brca2 -deficient groups. l The percentage of oocytes with misaligned chromosome in control and Brca2 -deficient groups. Statistical data were presented as mean percentage (mean ± SEM) of at least three independent experiments. Asterisk denotes statistical difference at a p < 0.05 level of significance

Article Snippet: Rabbit polyclonal anti-Brca2 antibody was purchased from Santa Cruz Biotechnology (Dallas, TA, USA; Cat#: sc-293185); mouse monoclonal anti-α-tubulin-fluorescein isothiocyanate (FITC) antibody was purchased from Sigma (St. Louis, MO, USA; Cat#: F2168); Rabbit monoclonal anti-γH2AX and anti-Gapdh antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA; Cat#: 9718, 2118); HRP-conjugated secondary antibodies were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA, USA); FITC-conjugated goat anti-rabbit IgG (H + L), TRITC-conjugated goat anti-rabbit IgG (H + L) and FITC-conjugated goat anti-mouse IgG (H + L) were purchased from Zhongshan Golden Bridge Biotechnology Co., LTD (Beijing, China).

Techniques: In Vivo, Control, Immunofluorescence, Fluorescence, In Vitro

Journal: Cell Death & Disease

Article Title: BRCA2 deficiency is a potential driver for human primary ovarian insufficiency

doi: 10.1038/s41419-019-1720-0

Figure Lengend Snippet: a Immunoblotting analysis of Brca2-HA protein expression in Brca2 -deficient oocytes. The blot was probed with anti-Brca2 antibody. The amount of Gapdh was used as an internal control. Two hundered germinal vesicle oocytes isolated from ovaries of 2-month-old mice were used for each lane of the blots. For each experiment, at least five mice of each genotype were used. b Representative images of MII oocytes matured in vitro from control ( Brca2 -proficient), Brca2 -deficient, and Brca2-rescued GV oocytes. Scale bar, 20 μm. c The rate of first polar body extrusion of control, Brca2 -deficient, and Brca2-rescued oocytes matured in vitro. d Representative images of spindle morphology and chromosome alignment in control, Brca2 -deficient, and Brca2-rescued oocytes after in vitro maturation. Oocytes were immunostained with anti-α-tubulin-FITC antibody to visualize spindles and counterstained with Hoechst to visualize chromosomes. Scale bar, 20 μm. ( e , f ) The rates of disorganized spindles and misaligned chromosomes in control, Brca2 -deficient and Brca2-rescued oocytes. g Immunofluorescence of γH2AX in control, Brca2 -deficient and Brca2-rescued oocytes. h Fluorescence intensity of γH2AX signal in control, Brca2 -deficient and Brca2-rescued oocytes were summarized. Statistical data were presented as mean percentage (mean ± SEM) of at least three independent experiments. Asterisk denotes statistical difference at a p < 0.05 level of significance

Article Snippet: Rabbit polyclonal anti-Brca2 antibody was purchased from Santa Cruz Biotechnology (Dallas, TA, USA; Cat#: sc-293185); mouse monoclonal anti-α-tubulin-fluorescein isothiocyanate (FITC) antibody was purchased from Sigma (St. Louis, MO, USA; Cat#: F2168); Rabbit monoclonal anti-γH2AX and anti-Gapdh antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA; Cat#: 9718, 2118); HRP-conjugated secondary antibodies were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA, USA); FITC-conjugated goat anti-rabbit IgG (H + L), TRITC-conjugated goat anti-rabbit IgG (H + L) and FITC-conjugated goat anti-mouse IgG (H + L) were purchased from Zhongshan Golden Bridge Biotechnology Co., LTD (Beijing, China).

Techniques: Western Blot, Expressing, Control, Isolation, In Vitro, Immunofluorescence, Fluorescence

Journal: Gynecologic Oncology Reports

Article Title: BRCA immunohistochemistry for screening of BRCA mutation in epithelial ovarian cancer patients

doi: 10.1016/j.gore.2020.100582

Figure Lengend Snippet: Performance of BRCA1 immunohistochemistry (IHC) and BRCA mutation status.

Article Snippet: Samples were stained and incubated for 60 min with BRCA1 mouse monoclonal antibody (Novus biological Inc., USA) and BRCA2 rabbit polyclonal antibody (Novus biological Inc., USA) at a dilution ratio of 1:100.

Techniques: Immunohistochemistry, Mutagenesis

Journal: Gynecologic Oncology Reports

Article Title: BRCA immunohistochemistry for screening of BRCA mutation in epithelial ovarian cancer patients

doi: 10.1016/j.gore.2020.100582

Figure Lengend Snippet: Performance of BRCA2 immunohistochemistry (IHC) and BRCA mutation status.

Article Snippet: Samples were stained and incubated for 60 min with BRCA1 mouse monoclonal antibody (Novus biological Inc., USA) and BRCA2 rabbit polyclonal antibody (Novus biological Inc., USA) at a dilution ratio of 1:100.

Techniques: Immunohistochemistry, Mutagenesis

Journal: Gynecologic Oncology Reports

Article Title: BRCA immunohistochemistry for screening of BRCA mutation in epithelial ovarian cancer patients

doi: 10.1016/j.gore.2020.100582

Figure Lengend Snippet: Performance of BRCA 1/2 immunohistochemistry (IHC) for detection of germline BRCA1/2 mutation.

Article Snippet: Samples were stained and incubated for 60 min with BRCA1 mouse monoclonal antibody (Novus biological Inc., USA) and BRCA2 rabbit polyclonal antibody (Novus biological Inc., USA) at a dilution ratio of 1:100.

Techniques: Immunohistochemistry, Mutagenesis

Journal: Med (New York, N.Y.)

Article Title: Inactivation of TACC2 epigenetically represses CDKN1A and confers sensitivity to CDK inhibitors.

doi: 10.1016/j.medj.2024.12.002

Figure Lengend Snippet: Figure 3. TACC2 interferes with the NuRD and CoREST corepressor complexes (A) SDS-PAGE and silver staining analysis of proteins immunoprecipitated using an anti-GFP antibody from EGFP-TACC2- or EGFP-overexpressing cells (left). The major interacting proteins are indicated on the right. (B) Schematic of N-terminal EGFP-tagged full-length TACC2 (FL) and the indicated deletion mutants (top) and IP coupled with immunoblot analysis of HEK293T cells transfected with the indicated constructs using the indicated antibodies (bottom). (C) IP of the endogenous TACC2 in ECA109 and KYSE30 cells followed by WB assays using the indicated proteins. (D) HEK293T cells were cotransfected with 3 mg each of vectors expressing FLAG-MTA1 and Myc-MBD3 or expressing FLAG-HMG20B and Myc-LSD1 together with 0, 3, and 6 mg of the vector expressing EGFP-TACC2 for 36 h. IP and WB were then performed with the indicated antibodies. (E) ECA109 cells were transfected with two TACC2 siRNAs and siNC for 48 h. IP and WB were performed with the indicated antibodies. (F) Representative immunofluorescence images of endogenous TACC2, MTA1, MBD3, or HMG20B in ECA109 cells. Green, TACC2; red, MTA1, MBD3, and HMG20B; blue, nuclei. Scale bars, 20 mm. (G) Representative immunofluorescence images of ECA109 cells transfected with TACC2 siRNAs or siNC for 48 h and stained with the indicated antibodies (left) and quantification of the nuclear localization percentage of MTA1, MBD3, or HMG20B (right). Scale bars, 20 mm. At least 100 cells were analyzed. Data are the mean ± SD of 3 independent experiments. Significance was measured by one-way ANOVA; *p < 0.05, **p < 0.01, ***p < 0.001. (H) Cytoplasmic and nuclear proteins from ECA109 cells transfected with TACC2 siRNAs or siNC for 48 h were separated and subjected to WB. (I) IHC analyses were performed with the indicated antibodies on serial sections of ESCC specimens (n = 45). Representative images are shown (left). Scale bars, 200 mm. Correlation between TACC2 levels and the cytoplasm localization of indicated proteins was determined (right). Two-tailed chi-square test; ***p < 0.001. See also Figure S5 and Table S3.

Article Snippet: After the retrieval solution had cooled to room temperature, sections were incubated with diluted anti-TACC2 antibody (rabbit polyclonal, 1:200; 11407-1-AP; Proteintech), anti-Ki67 antibody (rabbit polyclonal, 1:1000; 27309-1-AP; Proteintech), anti-Ki67 antibody (rabbit monoclonal, 1:200; #12202; CST), antiCD3 antibody (rabbit monoclonal, 1:100; ab5690; Abcam), anti-CD11b antibody (rabbit monoclonal, 1:100; ab133357; Abcam), anti-p21 antibody (rabbit monoclonal, 1:200; #2947; CST), anti-MTA1 antibody (rabbit monoclonal, 1:200; #5646; CST), antiMBD3 antibody (rabbit polyclonal, 1:200; 14258-1-AP; Proteintech), anti-HMG20B antibody (rabbit polyclonal, 1:200; 14582-1- AP; Proteintech), overnight at 4 C. Sections were then washed three times with phosphate-buffered saline containing Tween 20 (PBST; ZSGB-BIO) and incubated with goat anti-rabbit peroxidase-conjugated secondary antibodies (DAKO, Santa Clara) for 30min at 37 C. Sections were thenwashed three timeswith PBST and stainedwith 3,30-diaminobenzidine (DAB) for 2min to visualize the target protein.

Techniques: SDS Page, Silver Staining, Immunoprecipitation, Western Blot, Transfection, Construct, Expressing, Plasmid Preparation, Staining, Two Tailed Test