Figure 3
From: A p53-independent role for the MDM2 antagonist Nutlin-3 in DNA damage response initiation
Nutlin-3 leads to phosphorylation of several important DDR mediators. (A) HCT116p53+/+ cells were untreated (treated with DMSO only) (unt) or treated with 100 μM Etoposide (Eto) or 10 μM Nutlin-3 (Nut) for the times indicated before immunoblotting was used to analyse phosphorylation of p53 at Ser15, Ser20 and Ser37. Actin levels were used to assess equal loading. (B) HCT116p53+/+ cells were untreated (treated with DMSO only) (unt) or treated with 10 μM Nutlin-3 (Nut) or 100 μM of Etoposide (Eto) for 1 hour before the phosphorylation of ATM (Ser1981), BRCA1 (Ser1542) and CHK2 (Thr68) were analysed using immunoblotting. Actin levels were used to assess equal loading. (C) HCT116p53-/- cells were untreated (treated with DMSO only) (unt) or treated with 10 μM Nutlin-3 or 100 μM of Etoposide (Eto) for 1 hour before the phosphorylation of ATM (Ser1981) and BRCA1 (Ser1542) were analysed using immunoblotting. Actin levels were used to assess equal loading.