293 T cells were transfected with His-SAFB2, HA-ubiquitin, BRCA1, and BARD1 as indicated. Forty-eight hours after transfection, His-SAFB2 was isolated using nickel affinity chromatography under denaturing conditions. Ubiquitination of His-SAFB2 was analyzed by anti-HA immunoblotting (top), and His-SAFB2 was detected by anti-SAFB immunoblotting (bottom).