Total protein was extracted from the cells with RIPA lysis buffer (Beyotime Institute of Biotechnology) after transfection for 72 h and the concentration of the protein was determined with a Pierce™ BCA Protein assay kit (Peirce; Thermo Fisher Scientific, Inc.). Protein samples (15-30 μg/lane) were separated via 10% SDS-PAGE, and then separated proteins were transferred to PVDF membranes (EMD Millipore). The membranes were blocked with 5% BSA (Guangzhou Saiguo Biotech Co., Ltd.) for 1 h at room temperature. Subsequently, primary antibodies were added and incubated overnight at 4°C. Following washes with TBS with 0.05% Tween-20, the membranes were incubated with a secondary antibody [anti-mouse (cat. no. A0216) and anti-rabbit (cat. no. A0208) IgG (H+L) antibodies; 1:1,000; Beyotime Institute of Biotechnology] for 1 h at room temperature. Protein bands were visualized using an ECL kit (Shanghai Yeasen Biotechnology Co., Ltd.). The following primary antibodies were used: Anti-GMPS (1:1,000; cat. no. 14602; Cell Signaling Technology, Inc.), anti-P53 (1:1,000; cat. no. 2527; Cell Signaling Technology, Inc.), anti-phosphorylated (p)-P53 (1:1,000; cat. no. 2521; Cell Signaling Technology, Inc.), anti-Stat3 (1:1,000; cat. no. 9139; Cell Signaling Technology, Inc.), anti-p-Stat3 (1:1,000; cat. no. 9145; Cell Signaling Technology, Inc.) and anti-GAPDH (1:2,000; cat. no. 97166; Cell Signaling Technology, Inc.).

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