Primary antibody against trim59

The immunofluorescence was performed the same as the previous reports 13 (link), 14 (link). The frozen human ovarian cancer and the normal ovarian tissues were sectioned with a thickness of 10 μm. Primary antibody against TRIM59 (Abcam) were used overnight at 4°C. The cell nucleus was counterstained with 5 μg/ml DAPI (Beyotime). The intracellular distributions of the target proteins were analyzed and photographed using confocal fluorescent microscopy with excitation at 488 nm (for FITC) and 340 nm (for DAPI).

Western blotting analysis was performed according to the manufacturer’s protocols. Total proteins from both human tissues and cultured cells were extracted with lysis buffer, and the protein quality and quantity were detected by the Bradford dye-binding protein assay (Bio-Rad Laboratories, Hercules, CA, USA). A total of 50 μg of protein was loaded onto a 10% SDS polyacrylamide gel, after which PVDF membrane was used to transfer the proteins. The primary antibody against TRIM59 was purchased from Abcam (Cambridge, MA, USA), and primary antibody against GAPDH and secondary antibodies were commercially purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).