Journal:

Article Title: Modulation of DNA damage-induced apoptosis by cell adhesion is independently mediated by p53 and c-Abl

doi: 10.1073/pnas.1635435100

Figure Lengend Snippet: p53 and c-Abl contribute to araC-induced apoptosis. MEFs were left adhered to tissue culture plastic or detached and kept in suspension. At 2 h araC was added, and after 48 h caspase 3 activity was assayed. (A and B) WT and p53-/- MEFs, respectively, were analyzed without STI571. (C and D) The same cells were analyzed in the presence of 10 μM STI571. Values are means ± SEM from three independent experiments.

Article Snippet: Samples were assayed in lysis buffer supplemented with 100 mM NaCl and 10% glycerol by using the colorimetric substrate Asp-Glu-Val-Asp (DEVD)- p -nitoanilide with or without the caspase-3 inhibitor Ac-DEVD-CHO (Alexis Biochemicals, San Diego).

Techniques: Activity Assay

Journal:

Article Title: Modulation of DNA damage-induced apoptosis by cell adhesion is independently mediated by p53 and c-Abl

doi: 10.1073/pnas.1635435100

Figure Lengend Snippet: Analysis of p53-/- and c-Abl-/- cells. MEFs isolated from p53-/-/abl+/+ or p53-/-/abl-/- mice were left adhered to tissue culture plates or detached and kept in suspension. At 2 h, the indicated chemotherapeutic agent was added, and at 48 h caspase 3 activity or TUNEL stain was assayed. Values are means ± SEM from three independent experiments.

Article Snippet: Samples were assayed in lysis buffer supplemented with 100 mM NaCl and 10% glycerol by using the colorimetric substrate Asp-Glu-Val-Asp (DEVD)- p -nitoanilide with or without the caspase-3 inhibitor Ac-DEVD-CHO (Alexis Biochemicals, San Diego).

Techniques: Isolation, Activity Assay, TUNEL Assay, Staining

Journal:

Article Title: Modulation of DNA damage-induced apoptosis by cell adhesion is independently mediated by p53 and c-Abl

doi: 10.1073/pnas.1635435100

Figure Lengend Snippet: Role of p73. (A) Adherent or suspended cells were treated with araC or cisplatin (CDDP) as indicated, and p73 levels were assayed by Western blotting. A nonspecific band at 35 kDa was used as a loading control. (B) Cells were treated with araC or cisplatin in the presence of the indicated concentrations of STI571, then p73 was assayed as in A. (C) Apoptosis of p73-/- and WT MEFs after 48 h with or without 10 μM araC was compared by assaying caspase-3 activity. Both cell types are p53+/+. Values are means ± SEM (n = 3).

Article Snippet: Samples were assayed in lysis buffer supplemented with 100 mM NaCl and 10% glycerol by using the colorimetric substrate Asp-Glu-Val-Asp (DEVD)- p -nitoanilide with or without the caspase-3 inhibitor Ac-DEVD-CHO (Alexis Biochemicals, San Diego).

Techniques: Western Blot, Activity Assay

Journal:

Article Title: Modulation of DNA damage-induced apoptosis by cell adhesion is independently mediated by p53 and c-Abl

doi: 10.1073/pnas.1635435100

Figure Lengend Snippet: Cell type specificity. HT1080 fibrosarcoma, M21L melanoma, or C4-2 prostate carcinoma cells were left adherent on the dish (gray bars) or detached and kept in suspension (black bars). Cells were treated with the indicated concentrations of araC with or without 10 μM STI571 as indicated. Caspase-3 activity was assayed at 48 h. Values are means ± SEM from three independent experiments.

Article Snippet: Samples were assayed in lysis buffer supplemented with 100 mM NaCl and 10% glycerol by using the colorimetric substrate Asp-Glu-Val-Asp (DEVD)- p -nitoanilide with or without the caspase-3 inhibitor Ac-DEVD-CHO (Alexis Biochemicals, San Diego).

Techniques: Activity Assay