Primary antibodies against IKBa, pIKBa, p65, pp65, IKKa, and pIKKa were procured from Cell Signaling Technology (MA, USA).
The primary antibodies of rabbit anti-p16 were from Cell Signaling Technology, MA, USA, mouse anti-actin from Chemicon, and horseradish peroxidise-coupled second antibodies from Dako.
Rabbit anti-Keap1, anti-p62, anti-Akt (phosphor S473) monoclonal antibodies as well as anti-mTOR (phosphor S2448) polyclonal antibody were purchased from Cell Signaling Technology (MA, USA).
The membrane was immune-blotted with ERK1/2, phospho-ERK1/2, p38 MAPK and phospho-p38 MAPK (Cell signaling Technology, MA) to evaluate the phosphorylation of the compounds.
Anti-53BP1 (Bethyl laboratory Inc., Montgomery, TX), anti-JNK2 (D2) and anti-CDT1 (Santa Cruz Biotechnology, CA), anti-cleaved caspase 3 (Cell Signaling Technology, MA), anti-p53 (Imgenex, San diego, CA), anti-Rb, anti-p21Cip1 (BD Pharmingen, San Jose, CA), and mouse anti-GAPDH (Advanced ImmunoChemical Inc).
All antibodies were purchased from Cell Signaling Technology, MA, USA.
The cells were harvested for immunoprecipitation with ANTI-FLAG M2 Affinity Gel (Sigma, St . Louis MO) and were then subjected to the ChIP assay following the manufacturer's instruction of Chromatin immunoprecipitation kit (9002, Cell Signaling Technology, MA).
She is currently enrolled in an Education Technology, MA program at Teachers College, Columbia University.
Molecular weights standards were 'Biotinylated Protein Ladder Detection Pack' (Cell Signaling Technology, MA).
Additionally, protein extracts were analyzed by western-blotting using anti-caspase 3 and anti-activated caspase 3 antibodies (Cell Signaling Technology, MA).
Activation of caspase 9 was assayed by western blot using rabbit polyclonal antibody recognizing the cleaved/active variant of the protein (Cell Signaling Technology, MA, USA).
