Rad51

Total protein from the cell was collected using the ProteinExt^®Mammalian Total Protein Extraction Kit (TransGen Biotech, Beijing, China), following the manufacturer’s protocol. The concentration of the protein was measured using the Bradford protein assay kit (Novoprotein, Shanghai, China). The protein was resolved on 10% SDS-PAGE and then transferred to a PVDF membrane, followed by sealing of the sealing fluid. The membranes were incubated with the corresponding primary antibodies for 8 h at 4 °C and subsequently incubated with the secondary anti-bodies (Goat Anti-Rabbit IgG H&L (HRP), Zen bioscience, Chengdu, China) for 2 h. Then the bands were washed three times, and the hypersensitive chromogenic solution was developed using chemiluminescence in Touch Imager Pro, e-BLOT Life Science (Shanghai) Co., Ltd. (Shanghai, China). The primary antibodies PCNA, CDK2, and CDK4 were purchased from Zen bioscience (Chengdu, China). The primary antibodies PPARγ, CEBP/α, FABP4, SREBP1, and RAD51 were purchased from the Abclonal (Wuhan, China).

Samples were dissociated in RIPA lysis buffer containing a proteinase inhibitor cocktail. Once protein concentration was determined with the BCA Protein Assay Kit, total proteins were separated on SDS-PAGE gels and transferred onto the PVDF membranes. After blocking, the membranes were incubated overnight with primary antibodies at 4 °C. Primary antibodies were listed below: γH2AX (#2577S, Cell Signaling Technology), H2AX (#7631, Cell Signaling Technology), H3 (#17168-1-AP, Proteintech), POLE (#132100, Genetex), pTBK1 (#5483, Cell Signaling Technology), TBK1 (#BM4038, Boster), cGAS (#15102, Cell Signaling Technology), Ku70 (#A0883, ABclonal), Ku80 (#A5862, ABclonal), Lig4 (#A1743, ABclonal), RAD50 (#A3078, ABclonal), RAD51 (#A2829, ABclonal), BRCA1 (#A11034, ABclonal), NBS1 (#7703, ABclonal), p95/NBS1 (#ab23996, Abcam) EXO1 (#A1941, ABclonal), PARP1 (#A2432, ABclonal), ATR (#A7247, ABclonal), p-STING (#50907T, Cell Signaling Technology), STING (#ab92605, Abcam), β-Actin (#AC026, Abcam), and GAPDH (#ab9485, Abcam). Following three washes, the membranes were probed with HRP-conjugated secondary antibodies at room temperature (RT) for 1 h. The target proteins were viewed with an ECL Plus Kit (Abcam).