The protein was resolved by SDS/PAGE and blotted on nitrocellulose membranes (Bio-Rad, Richmond, CA, USA, Cat: 162-0115) as previously described 2 (link), 7 (link). The nitrocellulose membranes were incubated with specific primary antibodies overnight. After incubation with secondary antibodies, immunoreactive proteins were visualized using the Enhanced Chemiluminescent Substrate (Thermo Scientific, Pittsburgh, PA, USA, Cat: 34095).
Primary antibodies against cleaved caspase 3 (Cat: 9664, 1:500), p53 (Cat: 2524, 1:1000), phospho-p53 (Cat: 9286, 1:500), β-catenin (Cat: 9582, 1:1000), γ-H2AX (Cat: 9718, 1:1000), α-tubulin (Cat: 3873, 1:1000), β-actin (Cat: 3700, 1:1000), Histone H3 (Cat: 3638, 1:1000) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Cat: 8884, 1:1000); the phospho-mitogen-activated protein kinase (MAPK) Family Antibody Sampler Kit (Cat: 9910, 1:1000); and HRP-linked secondary antibodies were from Cell Signaling Technology (Beverly, MA, USA).
Primary antibodies against cleaved caspase 3 (Cat: 9664, 1:500), p53 (Cat: 2524, 1:1000), phospho-p53 (Cat: 9286, 1:500), β-catenin (Cat: 9582, 1:1000), γ-H2AX (Cat: 9718, 1:1000), α-tubulin (Cat: 3873, 1:1000), β-actin (Cat: 3700, 1:1000), Histone H3 (Cat: 3638, 1:1000) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Cat: 8884, 1:1000); the phospho-mitogen-activated protein kinase (MAPK) Family Antibody Sampler Kit (Cat: 9910, 1:1000); and HRP-linked secondary antibodies were from Cell Signaling Technology (Beverly, MA, USA).
