P53 antibody do 1 sc 126

p53 antibody (DO-1): sc-126 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). NF-κB p65 antibody (#8242), phospho-NF-κB p65 (Ser468) (#3039), Phospho-IKKα (Ser176)/IKKβ (Ser177) (C84E11) Rabbit mAb (#2078), E-Cadherin (24E10) (#3195), N-Cadherin (D4R1H) (#13116) and phospho-NF-κB p65 (Ser 536) (#3031) antibodies were obtained from Cell Signaling (Beverly, MA, USA). Fascin antibody (ab78599) was obtained from Abcam. The anti-β-actin antibody (AC-15) was purchased from Sigma-Aldrich (St. Louis, MO, USA).

MRC5 primary human fibroblasts were treated with 5 µM nutlin‐3. Five days later, cell lysate was prepared in HKMG buffer containing 10 mM Hepes pH 7.9, 100 mM KCl, 5 mM MgCl2, 10% glycerol, 0.5% NP‐40, 1 mM DTT, and protease inhibitors (Complete EDTA‐free, Roche) and was precleared with preequilibrated streptavidin‐coupled Dynabeads (Invitrogen). Annealed pairs of complementary oligonucleotides with the sense oligonucleotide biotinylated at the 5′ end (Sigma) were used for DNA pulldown. Oligonucleotide sequences are listed in Supporting Information Table S3. These DNA probes were incubated overnight at 4°C with the cell lysate. DNA‐bound proteins were then collected by incubation for 1 hr at 4°C with streptavidin‐coupled Dynabeads (Invitrogen), washed four times in HKMG buffer, and separated by SDS‐PAGE. p53 was detected by western blot (p53 antibody, DO‐1, sc‐126; Santa Cruz).