Ube2f

All antibodies were purchased from the following sources: NEDD8 (Abcam ab81264), DNA-PKcs (Thermo Fisher Scientific MA5-13238; Abcam ab32566; Proteintech 19983-1-AP), p-DNA-PKcs S2056(Abcam ab18192), Ku70 (Santa cruz sc-17789), Ku80(Santa cruz sc-9034), UBA3(Santa cruz sc-377352), UBE2M(Abcam ab109507), UBE2F(Abcam ab185234), Flag(Sigma F3156), γH2Ax(Millipore 05-636), HUWE1(Bethyl A300-486A-T). SFB-NEDD8, SFB-NEDD8ΔG, SFB-ub, HA-NEDD8, HA-NEDD8 K48R/K60R/NoK constructs were described previously (SFB-tag: S-protein tag, Flag epitope tag and streptavidin-binding peptide tag)^{13 (link)}. His-myc-tagged-NEDD8/NEDD8ΔG and NEDP1/NEDP1 C163S constructs were generated by PCR and cloned into pcDNA3.1-Myc-HisB. Flag-tagged-DNA-PKcs-F, M and FATC fragment were cloned into pcDNA3.1. Flag-tagged HUWE1 as previously described^{22 (link)} was gifted from professor Genze Shao (Peking University School of Basic Medical Sciences). His-Myc-tagged-DNA-PKcs-M fragment K to R mutants (1–8) were generated by PCR mutagenesis and then cloned into pcDNA3.1-Myc-HisB. The siRNAs were synthesized by GenePharma (Shanghai China) and used at 10 μm final concentration. siRNA as follows: UBA3(AGAGAGAGAUUAUGAGCAA), UBE2M-1 (CAGAGGUCCUGCAGAACAA), UBE2M-2 (GAUGAGGGCUUCUACAAGA), UBE2F-1 (GGAAUAAAGUGGAUGACUA), UBE2F-2 (CAACAUAAAUACAGCAAGA), HUWE1-1 (CAUUGGAAAGUGCGAGUUA), HUWE1-2 (CUGUGAGAGUGAUCGGGAA).

Antibodies specific to C-PARP (Cell Signaling Technology, 5625), Cullin1 (Abcam, 75817), Cullin2 (Abcam, 166917), Cullin3 (Cell Signaling Technology, 2759), Cullin4A (Cell Signaling Technology, 2699), Cullin4B (Proteintech, 12916-1-AP), Cullin5 (Abcam, 184177), NAE (Cell Signaling Technology, 14321), NOXA (Cell Signaling Technology, 14766), P27 (Cell Signaling Technology, 3686), RBX1 (Abcam, 133565), UBA3 (Abcam, 124726), UBE2F (Abcam, 12932), and UBE2M (Abcam, 109507) were purchased commercially.