Mouse primary anti p53

Cells and tissues were gently lysed with lysis buffer for 1 h on ice and then centrifuged at 15,000 g and 4 °C for 15 min, and the supernatant was collected. After the lysates were precleared with 50 μl of Protein A agarose (Sigma-Aldrich) for 2 h, the precleared lysates were incubated with 2 μg of each specific antibody overnight at 4 °C and then incubated with 50 μl of Protein A agarose for 4 h at 4 °C and were washed seven times. Immune complexes were eluted by boiling for 10 min at 95 °C in SDS sample buffer, followed by Western blotting with mouse primary anti-Myc (1:1000, Cell Signaling Technology) or mouse primary anti-p53 (1:1000, Santa Cruz Biotechnology) antibodies for cells, and rabbit primary anti-Chi3L1 (1:1000, abcam, Cambridge, MA) or mouse primary anti-p53 (1:1000, Santa Cruz Biotechnology) antibodies for tissues.