Journal: Biomarker Insights

Article Title: mRNA Expression and Methylation of the RAD51 , ATM , ATR , BRCA1 , and BRCA2 Genes in Gastric Adenocarcinoma

doi: 10.1177/11772719231225206

Figure Lengend Snippet: Cross-analysis of the methylation of RAD51 and BRCA1 genes in tumor and non-tumor mucosa, considering the 37 cases with paired tumoral and non-tumoral samples.

Article Snippet: The following antibodies were used: ATM (St John’s Laboratory, London, UK, STJ97797, 1:400), ATR (Abcam, Cambridge, UK, ab178407, 1:100), BRCA1 (St John’s Laboratory, STJ113833, 1:300), RAD51 (St John’s Laboratory, STJ95330, 1:100), and BRCA2 (St John’s Laboratory, STJ91885, 1:100).

Techniques: Methylation

Journal: Biomarker Insights

Article Title: mRNA Expression and Methylation of the RAD51 , ATM , ATR , BRCA1 , and BRCA2 Genes in Gastric Adenocarcinoma

doi: 10.1177/11772719231225206

Figure Lengend Snippet: RAD51 , ATR , BRCA1 , BRCA2 , and ATM mRNA expression in gastric adenocarcinoma and non-tumor gastric mucosa samples.

Article Snippet: The following antibodies were used: ATM (St John’s Laboratory, London, UK, STJ97797, 1:400), ATR (Abcam, Cambridge, UK, ab178407, 1:100), BRCA1 (St John’s Laboratory, STJ113833, 1:300), RAD51 (St John’s Laboratory, STJ95330, 1:100), and BRCA2 (St John’s Laboratory, STJ91885, 1:100).

Techniques: Expressing

Journal: Biomarker Insights

Article Title: mRNA Expression and Methylation of the RAD51 , ATM , ATR , BRCA1 , and BRCA2 Genes in Gastric Adenocarcinoma

doi: 10.1177/11772719231225206

Figure Lengend Snippet: Comparison of RAD51 , ATR , BRCA1 , BRCA2 , and ATM mRNA expression in tumor and non-tumor mucosa, considering the paired samples of the same individuals. The results are presented in box plots, using a logarithmic scale (+1 was added as a constant to allow the logarithmic representation of null values).

Article Snippet: The following antibodies were used: ATM (St John’s Laboratory, London, UK, STJ97797, 1:400), ATR (Abcam, Cambridge, UK, ab178407, 1:100), BRCA1 (St John’s Laboratory, STJ113833, 1:300), RAD51 (St John’s Laboratory, STJ95330, 1:100), and BRCA2 (St John’s Laboratory, STJ91885, 1:100).

Techniques: Comparison, Expressing

Journal: Biomarker Insights

Article Title: mRNA Expression and Methylation of the RAD51 , ATM , ATR , BRCA1 , and BRCA2 Genes in Gastric Adenocarcinoma

doi: 10.1177/11772719231225206

Figure Lengend Snippet: Comparison between RAD51 , ATR , BRCA1 , BRCA2 , and ATM mRNA expression in patients with previous neoadjuvant chemotherapy and in those without neoadjuvant chemotherapy, considering tumor samples and non-tumor mucosa samples.

Article Snippet: The following antibodies were used: ATM (St John’s Laboratory, London, UK, STJ97797, 1:400), ATR (Abcam, Cambridge, UK, ab178407, 1:100), BRCA1 (St John’s Laboratory, STJ113833, 1:300), RAD51 (St John’s Laboratory, STJ95330, 1:100), and BRCA2 (St John’s Laboratory, STJ91885, 1:100).

Techniques: Comparison, Expressing

Journal: Biomarker Insights

Article Title: mRNA Expression and Methylation of the RAD51 , ATM , ATR , BRCA1 , and BRCA2 Genes in Gastric Adenocarcinoma

doi: 10.1177/11772719231225206

Figure Lengend Snippet: Association between immunohistochemistry and mRNA levels in tumor samples.

Article Snippet: The following antibodies were used: ATM (St John’s Laboratory, London, UK, STJ97797, 1:400), ATR (Abcam, Cambridge, UK, ab178407, 1:100), BRCA1 (St John’s Laboratory, STJ113833, 1:300), RAD51 (St John’s Laboratory, STJ95330, 1:100), and BRCA2 (St John’s Laboratory, STJ91885, 1:100).

Techniques: Immunohistochemistry

Journal: Biomarker Insights

Article Title: mRNA Expression and Methylation of the RAD51 , ATM , ATR , BRCA1 , and BRCA2 Genes in Gastric Adenocarcinoma

doi: 10.1177/11772719231225206

Figure Lengend Snippet: Correlation between RAD51 , ATR , BRCA1 , BRCA2 , and ATM mRNA expression and histological type, histological grade, perineural invasion, and vascular invasion in tumor samples.

Article Snippet: The following antibodies were used: ATM (St John’s Laboratory, London, UK, STJ97797, 1:400), ATR (Abcam, Cambridge, UK, ab178407, 1:100), BRCA1 (St John’s Laboratory, STJ113833, 1:300), RAD51 (St John’s Laboratory, STJ95330, 1:100), and BRCA2 (St John’s Laboratory, STJ91885, 1:100).

Techniques: Expressing

Journal: Biomarker Insights

Article Title: mRNA Expression and Methylation of the RAD51 , ATM , ATR , BRCA1 , and BRCA2 Genes in Gastric Adenocarcinoma

doi: 10.1177/11772719231225206

Figure Lengend Snippet: Correlation between RAD51 , ATR , BRCA1 , BRCA2 , and ATM mRNA expression and depth of invasion (pT), regional lymph node metastasis, distant metastasis, and stage in tumor samples.

Article Snippet: The following antibodies were used: ATM (St John’s Laboratory, London, UK, STJ97797, 1:400), ATR (Abcam, Cambridge, UK, ab178407, 1:100), BRCA1 (St John’s Laboratory, STJ113833, 1:300), RAD51 (St John’s Laboratory, STJ95330, 1:100), and BRCA2 (St John’s Laboratory, STJ91885, 1:100).

Techniques: Expressing

Journal: Biomarker Insights

Article Title: mRNA Expression and Methylation of the RAD51 , ATM , ATR , BRCA1 , and BRCA2 Genes in Gastric Adenocarcinoma

doi: 10.1177/11772719231225206

Figure Lengend Snippet: Comparison between the mRNA levels in the tumors of patients who died and those who did not, considering the following groups: total population, excluding neoadjuvant therapy, with only surgical treatment, and with adjuvant or neoadjuvant treatment.

Article Snippet: The following antibodies were used: ATM (St John’s Laboratory, London, UK, STJ97797, 1:400), ATR (Abcam, Cambridge, UK, ab178407, 1:100), BRCA1 (St John’s Laboratory, STJ113833, 1:300), RAD51 (St John’s Laboratory, STJ95330, 1:100), and BRCA2 (St John’s Laboratory, STJ91885, 1:100).

Techniques: Comparison, Adjuvant

Journal: Biomarker Insights

Article Title: mRNA Expression and Methylation of the RAD51 , ATM , ATR , BRCA1 , and BRCA2 Genes in Gastric Adenocarcinoma

doi: 10.1177/11772719231225206

Figure Lengend Snippet: Kaplan-Meier curves comparing the overall survival of the 24 patients with tumor mRNA expression above the median with that of the 24 below the median, for RAD51 , ATR , BRCA1 , BRCA2 , and ATM .

Article Snippet: The following antibodies were used: ATM (St John’s Laboratory, London, UK, STJ97797, 1:400), ATR (Abcam, Cambridge, UK, ab178407, 1:100), BRCA1 (St John’s Laboratory, STJ113833, 1:300), RAD51 (St John’s Laboratory, STJ95330, 1:100), and BRCA2 (St John’s Laboratory, STJ91885, 1:100).

Techniques: Expressing

Journal: PLoS Genetics

Article Title: Elevated temperature increases meiotic crossover frequency via the interfering (Type I) pathway in Arabidopsis thaliana

doi: 10.1371/journal.pgen.1007384

Figure Lengend Snippet: (A, B) γH2Ax foci counting. (A) Pollen mother cells containing chromosomes (DAPI, blue) and γH2Ax foci (red). (B) Boxplots of γH2Ax foci at 20˚C and 28˚C; mean shown as white circle. (C, D) RAD51 foci counting. (C) Pollen mother cells containing chromosomes (DAPI, blue) and RAD51 foci (red). (D) Boxplots of RAD51 foci at 20°C and 28°C; mean shown as white circle. p-values are indicated as follows: ns = p > 0.5, * = p ≤ 0.05, ** = p ≤ 0.01. Scale bars represent 5 μm.

Article Snippet: The slides were incubated overnight with primary antibodies diluted 1:200 (RAD51, γH2AX, MLH1) in blocking buffer (goat serum, AR0009, Bosterbio) at 4°C and then at 37°C for 60 min with secondary antibody (1:1000, Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 555, catalog # A-21428, Thermo Fisher Scientific).

Techniques:

Journal: bioRxiv

Article Title: Fork Slowing and Reversal as an Adaptive Response to Chronic ATR Inhibition

doi: 10.1101/2021.05.18.444697

Figure Lengend Snippet: (A) RAD51 quantification relative to the Western blot shown in . (B) Immunoblot analysis, and relative quantification, of U-2OS cells treated with the indicated combination of AZD6738 and B02. (C) U-2OS and MDA-MB-231 were treated with the indicated combination of ATRi (AZD6738, 0.4µM) and RAD51i (B02, 10µM) for 5 days. For HCT116-TetON-shATR, cells were treated with the indicated combination of doxycycline and RAD51i (B02, 10µM) for five days. On day five, cells were harvested, and cell viability was assessed by Trypan blue staining. A representative image of was obtained by crystal violet staining (see the STAR Methods). Plotted results are the mean of two independent experiments ±SD. ** p<0.1, *** p<0.01 One-way Anova test. (D) Immunoblot analysis of HCT116-TetON-shATR cells treated for five days with or without doxycycline. (E) Immunoblot analysis of U-2OS cells treated with the indicated combination of AZD6738 (0.4µM), B02 (10µM), and HU (1mM). (F) HCT116 cells were treated as described in . The DNA fiber protocol was done following the conditions as in . 400 individual fibers for each condition were scored. Dot plot and median of total tract length are shown (n=2). **** p<0.001 One-way Anova test.

Article Snippet: The following primary antibodies were used: anti-β-Actin (Thermo Fisher Scientific; #MA1-140), anti-pDNA-PKcs-S2056 (Thermo Fisher Scientific; #PA5-78130), anti-DNA-PKcs (Bethyl Laboratories; #A300-516A), anti-pRPA2-S4/8 (Bethyl Laboratories; #A300-245A), anti-RAD51 (Merck Millipore; #PC130), anti-pKAP1-S824 (Bethyl Laboratories; #A300-767A), anti-pATM-S1981 (Cell Signaling Technology; #13050S), anti-KAP1 (Bethyl Laboratories; #A300-274A), anti-pCHK1-S345 (Cell Signaling Technology; #2341), anti-CHK1 (Santa Cruz Biotechnology; #sc-8408), anti-pH2AX-S139 (Bethyl Laboratories; A300-081A), anti-BRCA1 (Cell Signaling Technology; #9010), anti-CTIP (Bethyl Laboratories; #A300-488A), anti-BLM (Bethyl Laboratories; #A300-110A), anti-V5 (Thermo Fisher Scientific; #R960-25).

Techniques: Western Blot, Staining

Journal: Oncology Reports

Article Title: Antitumor effects and mechanisms of olaparib in combination with carboplatin and BKM120 on human triple-negative breast cancer cells

doi: 10.3892/or.2018.6716

Figure Lengend Snippet: Combination of OLA, CBP and BKM promotes DNA damage in triple-negative breast cancer cells. (A) Cells (CAL51) were treated with vehicle (CON), 1.2 µM OLA, 0.5 µM CBP, 0.5 µM BKM or a combination, for 48 h. RAD51 and γ-H2AX expression was visualized by immunofluorescence using primary specific antibodies and Alexa Fluor 488-conjugated secondary antibodies. Nuclei were stained with DAPI. The percentage of RAD51 and γ-H2AX positive cells (≥5 foci) and the number of RAD51 and γ-H2AX foci/cell were determined by counting at least 100 cells from each sample. Images were captured at ×400 magnification.. Data are presented as the mean ± standard deviation of three independent experiments. *P>0.05. BKM, NVP-BKM120; CBP, carboplatin; CON, control (vehicle); OLA, olaparib; RAD51, DNA repair protein RAD51 homolog 1. Combination of OLA, CBP and BKM promotes DNA damage in triple-negative breast cancer cells. (B) Cells (MDA-MB-231) were treated with vehicle (CON), 1.2 µM OLA, 0.5 µM CBP, 0.5 µM BKM or a combination, for 48 h. RAD51 and γ-H2AX expression was visualized by immunofluorescence using primary specific antibodies and Alexa Fluor 488-conjugated secondary antibodies. Nuclei were stained with DAPI. The percentage of RAD51 and γ-H2AX positive cells (≥5 foci) and the number of RAD51 and γ-H2AX foci/cell were determined by counting at least 100 cells from each sample. Images were captured at ×400 magnification.. Data are presented as the mean ± standard deviation of three independent experiments. *P>0.05. BKM, NVP-BKM120; CBP, carboplatin; CON, control (vehicle); OLA, olaparib; RAD51, DNA repair protein RAD51 homolog 1.

Article Snippet: Cells were incubated with the following antibodies for 12 h at 4°C: Anti-BRCA1 (cat. no. YT0519), anti-53BP1 (cat. no. YT0024) and anti-DNA repair protein RAD51 homolog 1 (RAD51; cat. no. YT3967) (all ImmunoWay Biotechnology Company), and γ-H2AX (Ser139; cat. no. 9718; Cell Signaling Technology, Inc.).

Techniques: Expressing, Immunofluorescence, Staining, Standard Deviation, Control

Journal:

Article Title: Increased ionizing radiation sensitivity and genomic instability in the absence of histone H2AX

doi: 10.1073/pnas.122228699

Figure Lengend Snippet: Formation of IR-induced BRCA1 and RAD51 foci in H2AXΔ/Δ vs. H2AXFlox/Δ ES cells. Immunofluorescence of H2AXFlox/Δ and H2AXΔ/Δ cells unirradiated (A) or 6 h postirradiation with 20 Gy (B) to visualize foci of BRCA1 (rhodamine, red) and RAD51 (FITC, green). Nuclei appear blue (Topro-3). In the merged images, overlapping foci appear yellow.

Article Snippet: Cells were stained with phospho-H2AX-specific polyclonal (1 μg/ml), mouse BRCA1-specific monoclonal (1:5 dilution), and RAD51 polyclonal (Oncogene Science; 1:5 dilution) antibodies, and rhodamine- and FITC-conjugated secondary antibodies (Jackson ImmunoResearch; 1:50 dilution).

Techniques: Immunofluorescence