Establishment of RNF5/185 DKO 293MSR cells. (A and B) Schematic representation of the RNF5 (A) and RNF185 (B) -targeting gRNA sequences. Arrows indicate primer positions. PAM, protospacer adjacent motif. The locations of each start codon, stop codon and the catalytic cysteine residues of RNF5 (C42) and RNF185 (C39, C42) are also indicated. The sequences analyzed for each KO cell line are shown along with the deleted sequences.