![Figure 4. Suz12 is required for B- and T-lymphopoiesis. Representative plots of flow cytometric analysis of BM from approximately 8- to 10-weeks old Suz12+/+/Rag1CreKI, Suz12fl/+/Rag1CreKI, and Suz12fl/fl/Rag1CreKI mice. Numbers represent the average proportions of (A) hematopoietic stem and progenitor cells (lineage− Sca-1+ c-Kit+ [LSK]), lymphoid-primed multipotent progenitors ([LMPP], LSK Flt3hi), and common lymphoid progenitors ([CLPs], lineage− Sca-1+ c-Kitint Flt3+ IL7Rα+) populations, and (B) total B cells (B220+ CD19+), pre-B + pro-B (B220+ IgM-), immature B ([Imm. b] B220lo IgM+), recirculating B ([Recirc. B] B220hi IgM+), and pre-proB (NK1.1− CD11c− CD19− B220+ CD43+) populations. Data represent mean ± standard deviation, from approximately 7-12 mice per genotype. (C) Flow cytometric analysis of thymus from approximately 8- to 10-week-old Suz12+/+/Rag1CreKI, Suz12fl/+/Rag1CreKI, and Suz12fl/fl/Rag1CreKI mice. The numbers represent the proportions of cells gated from the indicated populations. Data represent mean ± standard deviation from approximately 7-12 mice per genotype. (D) Competitive BM transplantation with equal numbers of BM cells from 8-week-old Suz12+/+/Rag1CreKI or Suz12fl/fl/Rag1CreKI mice (test; CD45.2) and Suz12+/+ mice (competitor; CD45.1) transplanted into lethally irradiated CD45.1 recipient mice. Test donor-derived (CD45.2+) contribution to specific cell types was assessed in the BM (top panel), thymus (middle panel), and peripheral blood (bottom panel) 8 weeks after transplantation. Gating strategies for BM: LSK, lymphoid-primed multipotent progenitors (LMPP), common lymphoid progenitor (CLP), pre-proB, immature, and recirculating B-cell populations are defined as described above, pro-B (B220+ CD19+ c-Kit+ IgM−), pre-B (B220+ CD19+ CD25+ c-Kit− IgM−); thymus: double negative [DN] (CD4− CD8−), DN1 (DN CD44+ CD25−), DN2 (DN CD44+ CD25+), DN3 (DN CD44− CD25+), DN4 (DN CD44− CD25−), DP (CD4+ CD8+); blood: total test donor-derived (CD45.2+), B cells (CD19+), T cells (CD4+ and CD8+), myeloid cells ([M] Mac-1+). Data represent mean ± standard deviation from (n = 3 donor) mice. Each donor marrow is transplanted into 3 recipients per test, and the averaged mean from each test is used to calculate the final mean. A two-tailed Student t test was performed to test statistical significance between genotypes. *P < .05; **P < .005; ***P < .001.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/126/2/10.1182_blood-2014-12-615898/4/167f4.jpeg?Expires=1749186185&Signature=SrxHKmhGiXPc6c67iKjEbPMvgqx334Fw5B-ZjokRPDEeUAvoGqNyftjBk73gUsuUyc7hA1D29CjNwnkVHiQZn97RlELHz1O5GSaB7Bum0-AxH3ky3UChMTyFCeH9EuZHUkhuvKa3StWD1BfUaCXXobZY7a2yTzkr2oBslMRyU4rj~edCHFILR3clW5cUB35pmZtSkcFOCZh2YRUq18m-wKwROQ0jXLwy6LF~yktv~wT-CwJ0U~9Ojda7DmiqDwPYnc1OI9BNacJDadfxHAwHQh0nk9vTjsbOfZEJnNce~2Yav7U88MPK72VsWJ1pcrOgWbJsJywuY1HHtqxKJ9O06w__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Suz12 is required for B- and T-lymphopoiesis. Representative plots of flow cytometric analysis of BM from approximately 8- to 10-weeks old Suz12+/+/Rag1CreKI, Suz12fl/+/Rag1CreKI, and Suz12fl/fl/Rag1CreKI mice. Numbers represent the average proportions of (A) hematopoietic stem and progenitor cells (lineage− Sca-1+ c-Kit+ [LSK]), lymphoid-primed multipotent progenitors ([LMPP], LSK Flt3hi), and common lymphoid progenitors ([CLPs], lineage− Sca-1+ c-Kitint Flt3+ IL7Rα+) populations, and (B) total B cells (B220+ CD19+), pre-B + pro-B (B220+ IgM-), immature B ([Imm. b] B220lo IgM+), recirculating B ([Recirc. B] B220hi IgM+), and pre-proB (NK1.1− CD11c− CD19− B220+ CD43+) populations. Data represent mean ± standard deviation, from approximately 7-12 mice per genotype. (C) Flow cytometric analysis of thymus from approximately 8- to 10-week-old Suz12+/+/Rag1CreKI, Suz12fl/+/Rag1CreKI, and Suz12fl/fl/Rag1CreKI mice. The numbers represent the proportions of cells gated from the indicated populations. Data represent mean ± standard deviation from approximately 7-12 mice per genotype. (D) Competitive BM transplantation with equal numbers of BM cells from 8-week-old Suz12+/+/Rag1CreKI or Suz12fl/fl/Rag1CreKI mice (test; CD45.2) and Suz12+/+ mice (competitor; CD45.1) transplanted into lethally irradiated CD45.1 recipient mice. Test donor-derived (CD45.2+) contribution to specific cell types was assessed in the BM (top panel), thymus (middle panel), and peripheral blood (bottom panel) 8 weeks after transplantation. Gating strategies for BM: LSK, lymphoid-primed multipotent progenitors (LMPP), common lymphoid progenitor (CLP), pre-proB, immature, and recirculating B-cell populations are defined as described above, pro-B (B220+ CD19+ c-Kit+ IgM−), pre-B (B220+ CD19+ CD25+ c-Kit− IgM−); thymus: double negative [DN] (CD4− CD8−), DN1 (DN CD44+ CD25−), DN2 (DN CD44+ CD25+), DN3 (DN CD44− CD25+), DN4 (DN CD44− CD25−), DP (CD4+ CD8+); blood: total test donor-derived (CD45.2+), B cells (CD19+), T cells (CD4+ and CD8+), myeloid cells ([M] Mac-1+). Data represent mean ± standard deviation from (n = 3 donor) mice. Each donor marrow is transplanted into 3 recipients per test, and the averaged mean from each test is used to calculate the final mean. A two-tailed Student t test was performed to test statistical significance between genotypes. *P < .05; **P < .005; ***P < .001.
