brca2  (Bethyl)

Journal: Nucleic Acids Research

Article Title: PARP1 and PARP2 are dispensable for DNA repair by microhomology-mediated end-joining at double-ended DSBs

doi: 10.1093/nar/gkaf1437

Figure Lengend Snippet: PARPi-dependent MMEJ increase depends on HR. ( A ) MMEJ quantification using the reporter and experimental timeline from Fig. and in HT1080 cells following treatment with olaparib (5 µM), DNA-PKcsi (NU7441, 1 µM), or both (combo). Values are normalized to DMSO. Immunoblot of 53BP1 and histone H3 in parental HT1080 cells and isogenic TP53BP1 -KO cells. ( C ) MMEJ quantification in HT1080 parental cells and indicated isogenic 53BP1 -KO cells. Values are normalized to wild-type DMSO. Drug used: olaparib (5 µM). ( D ) Immunoblot of BRCA2 and actin in parental HT1080 cells and two isogenic BRCA2 -KO clonal lines. ( E ) MMEJ quantification in HT1080 parental cells and BRCA2 -KO clones. Values are normalized to wild-type DMSO. Drug used: olaparib (5 µM). Statistical analyses for panels (A), (C), and (E). Data represent three independent experiments, each the average of three technical replicates. Data are mean ± SEM. Statistical test, one way ANOVA with multiple comparison correction. ns: nonsignificant, * P <.05, ** P <.01, **** P <.0001.

Article Snippet: Antibodies used for immunoblotting: PARP1 (diluted 1:2000, Cell Signaling Technology #9542), PARP2 (diluted 1:2000, Active Motif #39744, Antibody has been discontinued), β-Actin (diluted 1:5000, Cell Signaling Technology #8457), WRN (diluted 1:2000, Cell Signaling Technology #4666, RRID:AB_10692114 ), EXO1 (diluted 1:1000, Cell Signaling Technology #63862), PAR (diluted 1:2000, Cell Signaling Technology #87733), 53BP1 (diluted 1:2000, Novus Biologicals #NB100-304), H3 (diluted 1:5000, Cell Signaling Technology #9715), anti-mouse IgG, HRP-linked (diluted 1:5000, Cell Signaling #7076), anti-rabbit IgG, HRP-linked (diluted 1:5000, Cell Signaling #7074), BRCA2 (diluted 1:1000, Bethyl #A303-434A, RRID:AB_10952240 ).

Techniques: Western Blot, Clone Assay, Comparison

Journal: Cell Death & Disease

Article Title: Tumor-derived apolipoprotein E confers resistance to temozolomide in pancreatic neuroendocrine tumors

doi: 10.1038/s41419-025-08317-1

Figure Lengend Snippet: A GO functional enrichment analysis in TMZ-treated cells reveals the activation of various DNA damage pathways. B ssGSEA analysis indicates the score of DNA damage pathways in APOE (10 ng/ml)-treated cells based on transcriptomic data at different time points. C The expression of different genes related to the Homologous recombination (HR) pathway in APOE (10 ng/ml)-treated cells. D Based on GEPIA and TCGA data, the correlation between APOE expression and the HR genes BRCA1 and BRCA2 in pancreatic tissues. E Western blotting and RT-PCR experiments revealed that APOE enhances the expression of the HR genes BRCA1/2. F Knocking down APOE decreased expression of the HR genes BRCA1/2 induced by TMZ. G Comparison of various DNA damage repair pathway scores among the 10%FBS culture medium, 10% DLPS culture medium, and APOE (10 ng/ml)+10% DLPS culture medium, based on the transcriptomic data (48 h). H Comparison of functional enrichment analysis between the APOE(10 ng/ml)+10% DLPS culture medium and 10% DLPS culture medium group. I Comparison of various DNA damage repair pathway genes among the 10% FBS culture medium, 10% DLPS culture medium, and APOE(10 ng/ml)+10% DLPS culture medium cell. J Comparison of various DNA damage repair pathway genes among the 10% FBS culture medium, 10% DLPS culture medium, 10% FBS culture medium + TMZ, and 10% DLPS culture medium + TMZ cell. K Cell fluorescence response of APOE reduced the proportion of TMZ-induced DNA damage cells. Data in the graph are shown as the means ± SD from three independent experiment. * P < 0.05, ** P < 0.01, *** P < 0.001, ns: no significance.

Article Snippet: Antibodies were obtained from the following sources, unless indicated otherwise: BRCA1 (Abclonal, A11549 and A11034), BRCA2 (Ablonal, A2435), β-catenin (Proteintech, 51067-2-AP), γH2AX (Abcam, ab81299), and β-actin (Abclonal, AC006).

Techniques: Functional Assay, Activation Assay, Expressing, Homologous Recombination, Western Blot, Reverse Transcription Polymerase Chain Reaction, Comparison, Fluorescence

Journal: Cell Death & Disease

Article Title: Tumor-derived apolipoprotein E confers resistance to temozolomide in pancreatic neuroendocrine tumors

doi: 10.1038/s41419-025-08317-1

Figure Lengend Snippet: A The correlation analysis was determined by a two-tailed nonparametric Spearman correlation analysis between TCF/LEF in Wnt pathways with HR score or HR genes based on the transcriptomic data of cohort 2. B , C mRNA and protein expression of HR genes BCRA1/2 expression after knockdown of TCF3. D , E mRNA and protein expression of HR genes BRCA1/2 in cells expressing TCF3 plasmid. F Predicting three TCF3 binding site in human BRCA1and BRCA2 promoter through JASPAR Website, respectively. G The ChIP-PCR experiments were conducted to verify the binding ability of TCF3 to the three binding sites of BRCA1 and BRCA2, respectively. H The ChIP-PCR experiments investigated the impact of APOE on the transcriptional activation of TCF for BRCA1/2. I RT-qPCR analysis of BRCA1/2 expression in BON1 following treatment with ICG-001 and TCF3 plasmid in BON1. J mRNA expression of BRCA1/2 in shTCF3 BON1 with or without transfection of the CTNNB1 plasmid in BON1. Data in the graph are shown as the means ± SD from three independent experiment. * P < 0.05, ** P < 0.01, *** P < 0.001, ns: no significance.

Article Snippet: Antibodies were obtained from the following sources, unless indicated otherwise: BRCA1 (Abclonal, A11549 and A11034), BRCA2 (Ablonal, A2435), β-catenin (Proteintech, 51067-2-AP), γH2AX (Abcam, ab81299), and β-actin (Abclonal, AC006).

Techniques: Two Tailed Test, Expressing, Knockdown, Plasmid Preparation, Binding Assay, Activation Assay, Quantitative RT-PCR, Transfection