Defective homeostatic responses of F5 T cells in the absence of IKK2. F5 T cells from F5 Rag1 −/− huCD2 iCre+ R26R EYFP Ikk2 fx/fx (IKK2 ko) and huCD2 iCre- littermates (IKK2 wt) were labeled with CTV cell dye, mixed ∼1:1, and 2 × 10 6 total cells were transferred to either Rag1 −/− , Il15ra −/− Rag1 −/− , Il7 −/− Rag1 −/− or Ly5.1 F5 Rag1 −/− hosts for 14 d. ( A ) Histograms are of cell dye labeling in the indicated host by the indicted donor population at d7 and d14 after transfer. ( B ) Relative cell frequencies of YFP + and YFP − ). Graphs show ratio of YFP + :YFP − F5 T cells, normalized to input ratio at day 1 after transfer in the indicated hosts. ( C ) mRNA was isolated from sorted CD8 + TCR hi T cells from F5 Rag1 −/− Ikk2 fx /fx R26R EYFP huCD2 iCre and huCD2 iCre -ve littermate. Total mRNA was sequenced by a Illumina Genome Analyzer IIx. Following normalization reads were displayed as normalized reads per kilobase of exon per million reads (nRPKM). Bar charts show expression level (nRPKM) of the indicated genes in control F5 Rag1 −/− huCD2 iCre- R26R EYFP Ikk2 fx/fx samples (black bars; IKK2 WT) vs. F5 Rag1 −/− huCD2 iCre+ R26R EYFP Ikk2 fx/fx samples (red bars; IKK2 KO). Data are mean ± SD RPKM from triplicate biological replicates. ( D ) Graphs shows cell recovery from spleen, normalized to day 1, of IKK2 WT and IKK2 KO F5 T cells cotransferred to Il7 −/− Rag1 −/− hosts. Data are representative of five independent experiments.
{}