BCR sequence data

AID is required for SHM and antibody affinity maturation. To evaluate the effect of USP10 on AID-mediated SHM and affinity maturation, we first examined the BCR mutation frequency in NP-CGG-immunized USP10-BWT and USP10-BKO mice by antibody gene sequencing. After USP10-BWT and USP10-BKO mouse GC B cells were sorted by FACS, we sequenced the IgH VH186.2 germline, which encodes an NP-specific antibody, and the JH4 intronic region, which is irrelevant for affinity selection, thus, serves as a pure indicator of SHM efficiency. The overall mutation frequency of both JH4 and VH186.2 was substantially decreased in the absence of USP10 (Figure 3G), suggesting that SHM was impaired.