Journal: Journal of Clinical Oncology

Article Title: Detecting BRCA2 Protein Truncation in Tissue Biopsies to Identify Breast Cancers That Arise in BRCA2 Gene Mutation Carriers

doi: 10.1200/JCO.2008.20.5211

Figure Lengend Snippet: Dual C-terminal and N-terminal BRCA2 immunohistochemistry (IHC) of hereditary breast cancers (A and B, respectively) and of sporadic breast cancers (C and D, respectively). Magnification is (left) ×20 and (right) ×100 for all panels. The upper panel for each pair is C-terminal IHC, and the lower panel is N-terminal IHC. (A) Samples are from a BRCA2-mutant cancer with 7231del5. Top panel stained with C-terminal BRCA2 antibody, and second panel stained with N-terminal BRCA2 antibody. Only a lymphocyte in the top panel stains. (B) Samples are from a patient with a 9654delTT BRCA2 mutation at similar magnifications and similar staining. (C and D) The paired panels are from adjacent sections of the same sporadic breast cancer sample. Note the similar nuclear staining with both antibodies for the sporadic cancer samples.

Article Snippet: The N-terminal antibody used was from R&D systems MAB2476 (a commercially available mouse monoclonal antibody directed against an E coli –derived recombinant human BRCA2 protein that spans amino acids 1-200), which was used at 3 μg/mL.

Techniques: Immunohistochemistry, Mutagenesis, Staining

Journal: Journal of Clinical Oncology

Article Title: Detecting BRCA2 Protein Truncation in Tissue Biopsies to Identify Breast Cancers That Arise in BRCA2 Gene Mutation Carriers

doi: 10.1200/JCO.2008.20.5211

Figure Lengend Snippet: Immunohistochemistry with 575A15 C-terminal monoclonal antibody on normal breast epithelial lobules. Upper panels: untreated antibody. Middle panels: antibody mixed with excess of immunizing peptide (BRCA2 amino acids 3284 to 3294: TFVSPAAKAGG). Lower panels: antibody mixed with excess of control BRCA2 peptide (BRCA2 amino acids between 3300 and 3400, obtained from Abcam (Cambridge MA). Magnification is (left) ×20 and (right) ×100.

Article Snippet: The N-terminal antibody used was from R&D systems MAB2476 (a commercially available mouse monoclonal antibody directed against an E coli –derived recombinant human BRCA2 protein that spans amino acids 1-200), which was used at 3 μg/mL.

Techniques: Immunohistochemistry

Journal: Journal of Clinical Oncology

Article Title: Detecting BRCA2 Protein Truncation in Tissue Biopsies to Identify Breast Cancers That Arise in BRCA2 Gene Mutation Carriers

doi: 10.1200/JCO.2008.20.5211

Figure Lengend Snippet: MCF7 cells were cultured for 48 hours in 10% charcoal-stripped serum/phenol red–free DMEM and were treated with 10 nmol/L estrogen for 5 and 30 minutes and for 1, 2, 4, and 24 hours. Samples were blotted with the 575A15 BRCA2 C-terminal monoclonal antibody. The 440 kDa band is the only band on the blot.

Article Snippet: The N-terminal antibody used was from R&D systems MAB2476 (a commercially available mouse monoclonal antibody directed against an E coli –derived recombinant human BRCA2 protein that spans amino acids 1-200), which was used at 3 μg/mL.

Techniques: Cell Culture

Journal: Journal of Clinical Oncology

Article Title: Detecting BRCA2 Protein Truncation in Tissue Biopsies to Identify Breast Cancers That Arise in BRCA2 Gene Mutation Carriers

doi: 10.1200/JCO.2008.20.5211

Figure Lengend Snippet: BRCA2 Mutations Identified in Patients With Breast Cancer

Article Snippet: The N-terminal antibody used was from R&D systems MAB2476 (a commercially available mouse monoclonal antibody directed against an E coli –derived recombinant human BRCA2 protein that spans amino acids 1-200), which was used at 3 μg/mL.

Techniques: Mutagenesis

Journal: Frontiers in Oncology

Article Title: Prognostic value and experimental validation of atherosclerosis-derived pathogenic genes in colorectal cancer

doi: 10.3389/fonc.2025.1728087

Figure Lengend Snippet: Validation of the expression of 6 hub genes in CRC and AS by molecular biology experiments. (A-F) RT-qPCR validation of the differential expression of prognostic genes between normal and tumor tissues in CRC patients. (G) Western blot validation of the differential expression of 2 newly identified genes (HMMR and PALB2) between normal and tumor tissues in CRC. (H) Quantitative analysis of the relative grayscale values of Western blot bands for the 2 newly identified genes (HMMR and PALB2) in normal and tumor tissues of CRC. Data are presented as mean ± standard deviation (SD), with statistical significance indicated as * p < 0.05, *** p < 0.001, and **** p < 0.0001. ns indicates no statistical significance.

Article Snippet: The membrane was blocked with QuickBlockTM Blocking Buffer for Western Blot (Beyotime Biotechnology, Jiangsu, China) at room temperature for 1 h, washed with TBST, and incubated overnight at 4°C with primary antibodies: HMMR (1:2000, 15820-1-AP, Proteintech), PALB2 (1:2000, 14340-1-AP, Proteintech), and β-actin (1:10,000, 66009-1-Ig, Proteintech).

Techniques: Biomarker Discovery, Expressing, Quantitative RT-PCR, Quantitative Proteomics, Western Blot, Standard Deviation

Journal: Frontiers in Oncology

Article Title: Prognostic value and experimental validation of atherosclerosis-derived pathogenic genes in colorectal cancer

doi: 10.3389/fonc.2025.1728087

Figure Lengend Snippet: IHC staining validation. (A) The differential expression of PALB2 and HMMR in CRC tissues compared with normal tissues; (B) displays the differential expression of HMMR , PALB2 and PRR11 in AS tissues versus normal tissues. * p < 0.05.

Article Snippet: The membrane was blocked with QuickBlockTM Blocking Buffer for Western Blot (Beyotime Biotechnology, Jiangsu, China) at room temperature for 1 h, washed with TBST, and incubated overnight at 4°C with primary antibodies: HMMR (1:2000, 15820-1-AP, Proteintech), PALB2 (1:2000, 14340-1-AP, Proteintech), and β-actin (1:10,000, 66009-1-Ig, Proteintech).

Techniques: Immunohistochemistry, Biomarker Discovery, Quantitative Proteomics

Journal: Oncotarget

Article Title: Complementary genetic screens identify the E3 ubiquitin ligase CBLC, as a modifier of PARP inhibitor sensitivity.

doi: 10.18632/oncotarget.3628

Figure Lengend Snippet: Figure 2: CBLC gene silencing causes PARP inhibitor sensitivity. A. Clonogenic dose-response survival curves from MCF7 human breast cancer cells infected with shRNA expression constructs targeting either BRCA1 or CBLC. Transduced cells were subsequently exposed to olaparib for 14 days at which point cell colonies were counted. Two different shRNA expression constructs targeting CBLC, shCBLC_1 and shCBLC_2, were used.****ANOVA p value <0.0001 for the dose response curves in either shCBLC_1 or shCBLC_2 transduced cells vs. shCONTROL transduced cells. Data from shBRCA1 transduced cells is shown as the positive control. B. Bar chart illustrating CBLC RT-PCR data from MCF7 cells expressing CBLC cDNA and transduced with shRNA expression constructs. *Student’s t test p value <0.05 for CBLC expression compared to shCONTROL tranduced cells. C. and D. Olaparib dose-response survival curves from MCF7 (C) or HS578T (D) human breast cancer cells transfected with siRNA targeting either BRCA2 or CBLC. Cells were transfected with siRNA and 48 hours later exposed to olaparib for a subsequent six days.****ANOVA p value <0.0001 for the dose response curves in siCBLC transfected cells vs. siCONTROL transfected cells. Data from siBRCA2 transfected cells is shown as the positive control. E. Bar chart illustrating CBLC RT-PCR data from MCF7 cells expressing CBLC cDNA and transfected with siRNA. * Student’s t test p value <0.05 for CBLC expression compared to siCONTROL transfected cells. Where error bars are shown these represent the standard error of the mean (SEM) from three independent experiments.

Article Snippet: Each experiment was repeated twice. cell-based assays Cell lines were transfected with SMARTpool siRNAs (Dharmacon, GE Healthcare) targeting BRCA2 and CBLC or siCONTROL A (Santa Cruz Biotech) using RNAiMax (Invitrogen) transfection reagent.

Techniques: Infection, shRNA, Expressing, Construct, Positive Control, Reverse Transcription Polymerase Chain Reaction, Transduction, Transfection

Journal: Molecular cell

Article Title: Replication gaps are a key determinant of PARP inhibitor synthetic lethality with BRCA deficiency

doi: 10.1016/j.molcel.2021.06.011

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Antibodies for western blot analysis included anti-β-actin (Sigma), anti-FANCJ (E67), anti-BRCA1 (Cell Signaling Technology), anti-p21 (Cell Signaling Technology), anti-CHD4 (Abcam), anti-BRCA2 (Abcam), anti-53BP1 (Novus Biological), anti-RADX (CXorf57, Abcam), anti-RPA70/RPA1 (Cell Signaling Technology), anti-RPA32/RPA2 (Abcam), anti-FEN1 (Abcam), anti-PARP1 (Abcam), anti-XRCC1 (Abcam), anti-LIG1 (Santa Cruz), anti-LIG3 (GeneTex) and anti-H2B (Cell Signaling Technology), anti-Cleaved Caspase-3 (Cell Signaling Technology) and anti-PARP (Cleaved PARP1, Cell Signaling Technology).

Techniques: Recombinant, Electron Microscopy, Plasmid Preparation, Imaging, CCK-8 Assay, CRISPR, shRNA, Negative Control, Construct, Software, Transfection, DNA Extraction