Journal: Oncogene

Article Title: An anti-sense construct of full-length ATM cDNA imposes a radiosensitive phenotype on normal cells.

doi: 10.1038/sj.onc.1202007

Figure Lengend Snippet: Figure 2 Expression of ATM protein in untransfected C3ABR cells and in pMAT2 stably transfected C3ABR cells. Cells were treated with 5 mM CdCl2 for 6, 9 and 12 h. Total cell extracts of 50 mg were immunoblotted with ATM3BA antibodies. Lane 1, untreated C3ABR; lane 2, C3ABR 6 h post-induction; lane 3, 9 h post-induction; lane 4, 12 h post-induction; lane 5, untreated pMAT2 transfected C3ABR; lane 6, transfected 6 h post- induction; lane 7, 9 h post-induction; lane 8, 12 h post-induction; lane 9 is L3 an A-T cell line that does not produce ATM protein and was used as a control. Equal loading was ensured by reblotting ®lters with anti-DNA-PKcs antibody (DPKI)

Article Snippet: Cells were treated or mocktreated with 5 mM CdCl2 for 6 h, irradiated with 20 Gy of radiation and incubated for 1 h. Cells were lysed in universal immunoprecipitation bu er and 1 mg of total cell lysate was immunoprecipitated with anti-SAPK antibody (SAPKb, Santa Cruz).

Techniques: Expressing, Stable Transfection, Transfection, Control

Journal: Oncogene

Article Title: An anti-sense construct of full-length ATM cDNA imposes a radiosensitive phenotype on normal cells.

doi: 10.1038/sj.onc.1202007

Figure Lengend Snippet: Figure 3 Survival of pMAT2 stably transfected C3ABR cells after g-radiation. Viability was determined as described in the Materials and methods. Untransfected C3ABR and AT1ABR cells were used as controls. CdCl2 treated (+), not treated (7). Each point represents the average of duplicate experiments. Bars, s.d. The radiation dose used was 4 Gy

Article Snippet: Cells were treated or mocktreated with 5 mM CdCl2 for 6 h, irradiated with 20 Gy of radiation and incubated for 1 h. Cells were lysed in universal immunoprecipitation bu er and 1 mg of total cell lysate was immunoprecipitated with anti-SAPK antibody (SAPKb, Santa Cruz).

Techniques: Stable Transfection, Transfection

Journal: Oncogene

Article Title: An anti-sense construct of full-length ATM cDNA imposes a radiosensitive phenotype on normal cells.

doi: 10.1038/sj.onc.1202007

Figure Lengend Snippet: Figure 4 Stabilization of p53 and induction of p21/WAF1 after exposure of cells to radiation. C3ABR cells and C3ABR transfected with pMAT2(+) were CdCl2 treated for 6 h (+), not treated (7) and exposed (+) to g-radiation (6 Gy) followed by 2 h incubation. p53 and p21/WAF1 were determined by immunoblotting with anti-p53 antibody (pAB 1801, Oncogene Science) or anti-WAF1 antibody (OP-64, Oncogene Science), respectively. PCNA represents a loading control

Article Snippet: Cells were treated or mocktreated with 5 mM CdCl2 for 6 h, irradiated with 20 Gy of radiation and incubated for 1 h. Cells were lysed in universal immunoprecipitation bu er and 1 mg of total cell lysate was immunoprecipitated with anti-SAPK antibody (SAPKb, Santa Cruz).

Techniques: Transfection, Incubation, Western Blot, Control

Journal: Oncogene

Article Title: An anti-sense construct of full-length ATM cDNA imposes a radiosensitive phenotype on normal cells.

doi: 10.1038/sj.onc.1202007

Figure Lengend Snippet: Figure 5 Examination of the rate of DNA synthesis in pMAT2 stably transfected C3ABR cells after g-radiation. The eect of radiation on the rate of DNA synthesis was measured after g- radiation as described in the Materials and methods. Untrans- fected C3ABR and AT1ABR cells were used as controls. CdCl2 treated (+), not treated (7). Each point represents the average of duplicate experiments. Bars, s.d.

Article Snippet: Cells were treated or mocktreated with 5 mM CdCl2 for 6 h, irradiated with 20 Gy of radiation and incubated for 1 h. Cells were lysed in universal immunoprecipitation bu er and 1 mg of total cell lysate was immunoprecipitated with anti-SAPK antibody (SAPKb, Santa Cruz).

Techniques: DNA Synthesis, Stable Transfection, Transfection

Journal: Oncogene

Article Title: An anti-sense construct of full-length ATM cDNA imposes a radiosensitive phenotype on normal cells.

doi: 10.1038/sj.onc.1202007

Figure Lengend Snippet: Figure 6 Extent of G2 phase delay at 24 h post-irradiation. Cells were incubated with 5 mM CdCl2 for 6 h, treated with 3 Gy of radiation, and pelleted. After resuspending in CdCl2 free growth media, the cells were further incubated for 24 h and applied to cell cycle analysis as previously described (Beamish and Lavin, 1994)

Article Snippet: Cells were treated or mocktreated with 5 mM CdCl2 for 6 h, irradiated with 20 Gy of radiation and incubated for 1 h. Cells were lysed in universal immunoprecipitation bu er and 1 mg of total cell lysate was immunoprecipitated with anti-SAPK antibody (SAPKb, Santa Cruz).

Techniques: Irradiation, Incubation, Cell Cycle Assay

Journal: Oncogene

Article Title: An anti-sense construct of full-length ATM cDNA imposes a radiosensitive phenotype on normal cells.

doi: 10.1038/sj.onc.1202007

Figure Lengend Snippet: Figure 7 Activation of SAPK by ionizing radiation. pMAT2 transfected C3ABR cells were induced or mock-induced for ATM anti-sense expression with 5 mM CdCl2 for 6 h. Cells were then treated with 20 Gy of radiation and harvested 1 h later. 1 mg of total lysate was immunoprecipitated with anti-SAPK antibody (SAPKb, Santa Cruz), and in vitro immune complex kinase assays were performed with GST-Jun (residues 1 ± 79) as substrate. The A-T cell line L3 was used as a negative control, since the mutation detected is only predicted to give rise to a 35 amino acid sized fragment

Article Snippet: Cells were treated or mocktreated with 5 mM CdCl2 for 6 h, irradiated with 20 Gy of radiation and incubated for 1 h. Cells were lysed in universal immunoprecipitation bu er and 1 mg of total cell lysate was immunoprecipitated with anti-SAPK antibody (SAPKb, Santa Cruz).

Techniques: Activation Assay, Transfection, Expressing, Immunoprecipitation, In Vitro, Immune Complex Kinase Assay, Negative Control, Mutagenesis