primary rabbit polyclonal antibodies against rad51  (Santa Cruz Biotechnology)

Journal: International Journal of Molecular Sciences

Article Title: The CD44high Subpopulation of Multifraction Irradiation-Surviving NSCLC Cells Exhibits Partial EMT-Program Activation and DNA Damage Response Depending on Their p53 Status

doi: 10.3390/ijms22052369

Figure Lengend Snippet: Kinetics of γH2AX and Rad51 foci changes in A549 and A549IR cells and H1299 and H1299IR cells after 2 Gy X-ray exposure. Representative immunofluorescent images of the irradiated cells showing Rad51 (green), γH2AX (red) foci and their colocolization (Merged). DAPI nuclear counterstaining is shown in blue ( a ). Comparative analysis of changes in the number of γH2AX foci in A549 and A549IR ( b ) and H1299 and H1299IR cells ( d ) after 2 Gy X-ray exposure; changes in the number of Rad51 foci in A549 and A549IR cells ( c ) and H1299 and H1299IR cells ( e ) after 2 Gy X-ray exposure. ↕ denotes significant differences between groups at p < 0.05. Data are means ± SD of three independent experiments.

Article Snippet: Cells were then incubated overnight at 4 ᵒC with primary rabbit polyclonal antibody against Rad51 (dilution 1:200, Merck Millipore, Burlington, VT, USA) and primary rabbit polyclonal antibody against DNA PKcs (phospho S2056) (dilution 1:200, Abcam, Cambridge, USA) or primary rabbit polyclonal antibody against LC3-I/II (dilution 1:200, Merck Millipore, Burlington, VT, USA).

Techniques: Irradiation

Journal: International Journal of Molecular Sciences

Article Title: The CD44high Subpopulation of Multifraction Irradiation-Surviving NSCLC Cells Exhibits Partial EMT-Program Activation and DNA Damage Response Depending on Their p53 Status

doi: 10.3390/ijms22052369

Figure Lengend Snippet: The proportion of Rad51 positive cells in the CD44low/CD166low and CD44high/CD166high populations of MFR-surviving and their parental NSCLC cells at 6 h and 24 h after exposure to 2 Gy and 5 Gy of X-ray: A549IR and A549 cells ( a ); H1299IR and H1299 cells ( b ). * p < 0.05. Data are means ± SEM of more than three independent experiments.

Article Snippet: Cells were then incubated overnight at 4 ᵒC with primary rabbit polyclonal antibody against Rad51 (dilution 1:200, Merck Millipore, Burlington, VT, USA) and primary rabbit polyclonal antibody against DNA PKcs (phospho S2056) (dilution 1:200, Abcam, Cambridge, USA) or primary rabbit polyclonal antibody against LC3-I/II (dilution 1:200, Merck Millipore, Burlington, VT, USA).

Techniques:

Journal: Oncology Reports

Article Title: M2 macrophages reduce the radiosensitivity of head and neck cancer by releasing HB-EGF

doi: 10.3892/or.2020.7628

Figure Lengend Snippet: In CAL27 cells, inhibition of EGFR by small molecule inhibitor results in reduced NHEJ repair capacity. (A) CAL27 and CAL27 with HB-EGF (50 ng/ml) were assessed by the western blot analysis with anti-EGFR and anti-p-EGFR antibodies. (B) EGFR and EGFR phosphorylation expression in CAL27 cells with or without HB-EGF after 4 Gy radiation. NC, CAL27 cells. (C) Representative images of the number of the p-DNA-PK foci per cell in 5 h after irradiation (4 Gy) in CAL27 cells with or without the EGFR inhibitor; magnification, ×200 (green, p-DNA-PK staining; blue, DAPI). (D) Representative images of the number of the RAD51 foci per cell in 5 h after irradiation (4 Gy) in CAL27 cells with or without the EGFR inhibitor; magnification ×200 (green, p-DNA-PK staining; blue, DAPI). *P<0.05. In CAL27 cells, inhibition of EGFR by small molecule inhibitor results in reduced NHEJ repair capacity. (E-a) Number of p-DNA-PK foci of NHEJ specific markers in CAL27 cells with or without EGFR inhibitors; (E-b) Number of RAD51 foci of HR specific markers in CAL27 cells with or without EGFR inhibitors. (F) IHC analyses of EGFR in cancer and normal tissues; magnification, ×400 (HPV + T, HPV-positive tumor tissues; HPV-T, HPV-negative tumor tissues; HPV + /HPV − N, head and neck normal tissues). (G) EGFR IHC score. HPV-positive cancer tissues (n=19), HPV-negative cases (n=33). (H) Expression of EGFR between HPV-positive (n=98) and HPV-negative (n=420) HNSCC in the TCGA database. Results are presented as the mean ± SD. *P<0.05. HPV, human papilloma virus; HNSCC, head and neck squamous cell carcinoma; EGFR, epidermal growth factor receptor; NHEJ, non-homologous end-joining; HB-EGF, heparin-binding epidermal growth factor; p-, phosphorylated; DNA-PK, DNA-dependent protein kinase; RAD51, RAD51 recombinase; IHC, immunohistochemistry; TCGA, The Cancer Genome Atlas.

Article Snippet: Samples were incubated with the γ H2A histone family member X (H2AX) mouse monoclonal antibody (product code ab26350; Abcam) at 1:500 dilution, primary rabbit polyclonal antibodies against RAD51 recombinase (RAD51; product code ab133534; Abcam) at 1:200, and phosphorylated (p)-DNA-dependent protein kinase (DNA-PK; phosphor S2056; product code ab124918; Abcam) at 1:200 and EGFR inhibitor gefitinib (ZD1839; cat. no. HY-50895; MedChemExpress) overnight at 4°C.

Techniques: Inhibition, Western Blot, Expressing, Irradiation, Staining, Non-Homologous End Joining, Binding Assay, Immunohistochemistry