Figure 1. Deletion of Ikk1 and Ikk2 blocks SP thymocyte development. Lymphoid organs from 8–12-wk-old Ikk1fx/fx Ikk2fx/fx R26REYFP huCD2iCre (IKKΔTCD2, n = 16) were analyzed by FACS and compared with huCD2iCre -ve (Cre−, n = 15) littermate as control. (A) Density plots are of CD44 versus CD25 by DN Lin− (CD4, CD8, TCRαβ, and CD5) thymocytes. Numbers indicate percentage in the corresponding quadrant. Bar charts are cell number of CD44hiCD25hi (DN2), CD44loCD25hi (DN3), and CD44lo CD25lo (DN4) DN thymocyte subsets. (B) Density plots are of TCR versus CD5. Numbers indicate percentage in the corresponding gate. Bar charts are cell number of CD5loTCRlo (DP1), CD5hiTCRint (DP2), and CD5intTCRhi (DP3) DP subsets. (C) Density plots are of CD4 versus CD8 expression by total live thymocytes and TCR versus HSA by CD4SP and CD8SP gated thymocytes. Bar charts show numbers of the indicated SP subset from IKKΔTCD2 mice (Cre+) and huCD2iCre -ve (Cre−) littermates. Numbers indicate fold difference in counts between Cre− and Cre+. (D) Density plots are of CD4 versus CD8 expression by total live thymocytes, TCR versus CD5 by DP thymocytes, and TCR versus HSA by CD4SP and CD8SP gated thymocytes from IKKΔTCD4 mice or Cre− littermates. (E) Density plots are CD25 versus CD44 by CD4+TCRhi gated lymph node cells and CD44 versus TCR by CD8+TCRhi gated lymph node cells from the indicated strains and show gates used to define naive (CD44lo CD25−), memory (CD44hi CD25−), and T reg (CD25+) cells among CD4+TCRhi cells, and naive (CD44lo) and memory (CD44hi) among CD8+TCRhi cells. Bar charts show total number of naive, memory, and T reg cells recovered from lymph node and spleen of IKKΔTCD2 mice (Cre+) and huCD2iCre -ve (Cre−) littermates. Numbers indicate fold difference in naive cell counts between Cre− and Cre+ hosts. (F) Histograms are of EYFP expression by naive, memory, and regulatory CD4+TCRhi T cells and naive and memory CD8+TCRhi T cells from IKKΔTCD2 mice and huCD2iCre Ikk1fx/WT Ikk2fx/WT R26REYFP littermate controls (WT). Numbers indicate percentage of IKKΔTCD2 cells (solid lines) expressing YFP. FACS data are representative of four or more experiments and cell number data are pooled from four experiments. ****, P < 0.0001.
Figure 1.

Deletion of Ikk1 and Ikk2 blocks SP thymocyte development. Lymphoid organs from 8–12-wk-old Ikk1fx/fx Ikk2fx/fx R26REYFP huCD2iCre (IKKΔTCD2, n = 16) were analyzed by FACS and compared with huCD2iCre -ve (Cre−, n = 15) littermate as control. (A) Density plots are of CD44 versus CD25 by DN Lin− (CD4, CD8, TCRαβ, and CD5) thymocytes. Numbers indicate percentage in the corresponding quadrant. Bar charts are cell number of CD44hiCD25hi (DN2), CD44loCD25hi (DN3), and CD44lo CD25lo (DN4) DN thymocyte subsets. (B) Density plots are of TCR versus CD5. Numbers indicate percentage in the corresponding gate. Bar charts are cell number of CD5loTCRlo (DP1), CD5hiTCRint (DP2), and CD5intTCRhi (DP3) DP subsets. (C) Density plots are of CD4 versus CD8 expression by total live thymocytes and TCR versus HSA by CD4SP and CD8SP gated thymocytes. Bar charts show numbers of the indicated SP subset from IKKΔTCD2 mice (Cre+) and huCD2iCre -ve (Cre−) littermates. Numbers indicate fold difference in counts between Cre− and Cre+. (D) Density plots are of CD4 versus CD8 expression by total live thymocytes, TCR versus CD5 by DP thymocytes, and TCR versus HSA by CD4SP and CD8SP gated thymocytes from IKKΔTCD4 mice or Cre− littermates. (E) Density plots are CD25 versus CD44 by CD4+TCRhi gated lymph node cells and CD44 versus TCR by CD8+TCRhi gated lymph node cells from the indicated strains and show gates used to define naive (CD44lo CD25−), memory (CD44hi CD25−), and T reg (CD25+) cells among CD4+TCRhi cells, and naive (CD44lo) and memory (CD44hi) among CD8+TCRhi cells. Bar charts show total number of naive, memory, and T reg cells recovered from lymph node and spleen of IKKΔTCD2 mice (Cre+) and huCD2iCre -ve (Cre−) littermates. Numbers indicate fold difference in naive cell counts between Cre− and Cre+ hosts. (F) Histograms are of EYFP expression by naive, memory, and regulatory CD4+TCRhi T cells and naive and memory CD8+TCRhi T cells from IKKΔTCD2 mice and huCD2iCre Ikk1fx/WT Ikk2fx/WT R26REYFP littermate controls (WT). Numbers indicate percentage of IKKΔTCD2 cells (solid lines) expressing YFP. FACS data are representative of four or more experiments and cell number data are pooled from four experiments. ****, P < 0.0001.

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