We next assessed GC formation and CSR after mouse immunization with sheep red blood cells (SRBC). and recirculating B cells from and control mice. Right, frequency quantification of each B cell subpopulation in (n = 9) and mice (n = 8). Two-tailed t-test, error bars represent SD.(TIF) pgen.1008960.s001.tif (938K) Dicoumarol GUID:?C4FE1E15-6763-4B1F-9336-60DDD4B17D5B S2 Fig: Mouse model for the conditional expression of AID in hematopoietic cells. FACS analysis of GFP reporter expression in bone marrow, spleen and thymus cells from (black empty line) and (grey shade). Bone marrow and spleen B cell subsets were gated as in S1BCS1C Fig. T cell subsets were gated as DN (CD4-CD8-), DN1 (CD4-CD8-CD44+CD25-), DP (CD4+CD8+), CD4+SP (CD4+CD8-), CD8+SP (CD4-CD8+) from thymus and naive-T (B220-CD3+) from spleen.(TIF) pgen.1008960.s002.tif (417K) GUID:?FBABA98A-7680-4660-9254-8BD15A242B5B S3 Fig: Staining controls for Fig 4D. Ki67 and AID immunohistochemistry Dicoumarol of spleen sections from SRBC immunized and mice. Magnification is 5x inset is GCN5 40x. Scale bars are 500m and 50m for 5x and 40x images respectively.(TIF) pgen.1008960.s003.tif (2.9M) GUID:?F1748186-A0FC-44B8-AC75-A7966FD3F98C S4 Fig: Immunohistochemistry analysis (left) and immunophenotype (right) of 10 tumors from mice. Magnification for Ki67 and AID IHQ images is 40x. Scale bar is 50m. Total live cells (left FACs panel) and B220+ gated cells (middle and right FACs panels) are shown.(TIF) pgen.1008960.s004.tif (6.3M) GUID:?CB360B0A-06BA-4D6F-A281-894E0C7B18D9 S5 Fig: Immunohistochemistry analysis (left) and immunophenotype (right) of 10 tumors from mice. Magnification for Ki67 and AID IHQ images is 40x. Scale bar is 50m. Total live cells (left FACs panel) and B220+ gated cells (middle Dicoumarol and right FACs panels) are shown.(TIF) pgen.1008960.s005.tif (6.5M) GUID:?6CDAE6BD-BD74-4604-B2AC-2D6A858BE343 S6 Fig: Exome sequencing of tumors. (A) Total number of SNVs and INDELS identified in each of the 15 tumors analyzed. (B) Average proportion of SNVs and INDELs found in and tumors. (C) CNV analysis of tumors from WES data. Heatmap of the genomic distribution of CNV alterations in and tumors. Upper panel depicts number of CNV regions per tumor, with colors encoding copy number gain (red) or loss (grey). (D) Number of translocations identified in and tumors by Manta analysis of WES data (two-tailed t-test, p = 0.852; tumor vs healthy tissue, see methods for details).(TIF) pgen.1008960.s006.tif (925K) GUID:?5C556E67-A2C1-429C-BDF4-338EDC2B0559 S1 Table: Whole Exome Sequencing (WES) metrics of the 16 samples analyzed. (XLSX) pgen.1008960.s007.xlsx (11K) GUID:?9A3E837C-8971-492E-932E-4CA1309132D4 S2 Table: Tumors samples analyzed by WES. (XLSX) pgen.1008960.s008.xlsx (13K) GUID:?A333CFB3-7293-4912-90B6-470C4CE6D3D1 S3 Table: Variants identified in AID KI UNG KO and AID WT UNG KO tumors. (XLSX) pgen.1008960.s009.xlsx (6.1M) GUID:?76E361B5-8A36-4325-BC8C-156EBFF5BB5E S4 Table: Traslocation analysis of AID KI UNG KO and AID WT UNG KO tumors. (XLSX) pgen.1008960.s010.xlsx (35K) GUID:?646956A0-0599-4E0B-B4AD-97E378AFB214 S5 Table: List of cancer-related genes mutated in AID KI UNG KO and/or AID WT UNG KO tumors. (XLSX) pgen.1008960.s011.xlsx (28K) GUID:?3F663355-5016-4D73-AC89-AA860E71F656 S1 Appendix: Mutation analysis of PCR-seq data at the IgH S region from GC and na?ve B cells. (XLSX) pgen.1008960.s012.xlsx (545K) GUID:?E417AF38-2514-4F66-9B30-FC2AA412D716 Attachment: Submitted filename: -MMR deficient- mice show different alterations in SHM and CSR [11C18]. Likewise, double deficient mice only retain transition mutations at C/G, directly emerging from the replication of U:G mismatches, and are devoid of other SHM footprints, as well as of CSR [19,20]. Early studies showed that several lymphoma-associated oncogenes bore mutations with the hallmark of SHM [21C23], and the molecular contribution of AID activity to off-target SHM was directly shown in mice later [20,24], indicating that a relatively high proportion of genes can be targets of AID mutagenic activity. Moreover, AID can trigger chromosome translocations involving and translocations are abolished in the absence of UNG [25], suggesting that UNG activity is required for the processing of AID deaminations into the DNA double strand break preceding the chromosome translocation joining. On the other hand, SHM frequency is increased in the combined absence of UNG and MSH2, indicating that BER and MMR contribute together to the faithful repair of AID-induced deaminations [4,20,24,30]. In line Dicoumarol with these observations, UNG deficient mice develop B cell lymphomas [31], indicating a tumor suppressor role for UNG. However, the contribution of UNG to lymphomagenesis is intricate and not completely understood. On one hand, UNG deficiency protects against the development of BCL6-driven DLBCL.