Bcl-2, IL-7Rα chain, and Bim levels during deletion and immunity. A total of 2 × 10⁶ CFSE-labeled OT-I cells were injected intravenously into either RIP-OVA^hi mice or OCS/LPS primed C57BL/6 mice (OCS/LPS). Sixty hours after transfer, the proliferating cells within the sacral and pancreatic lymph nodes (RIP-OVA^hi) or spleen (OCS/LPS) were stained and analyzed by flow cytometry. (A) Intracellular Bcl-2 staining showing dot plots (left) and quantitated mean fluorescence intensity (MFI; right). MFI graph shows Bcl-2 MFI values minus background, which was determined independently for each cell division. Error bars represent SEM. Representative data are shown from 1 of 3 independent experiments, with 3 mice per group per experiment. (B) Bim real-time PCR performed on array cRNA. The bar graphs show changes in gene expression relative to naive and represent mean data ± SEM from the duplicate array samples (with the exception of the Rag sample, for which only a single replicate was analyzed). Naive indicates naive cells; Tol, cells undergoing deletion; Imm, primed cells; and Rag, cells undergoing lymphopenia-induced proliferation. (C) Same as in panel A, except that cells were stained intracellularly for Bim. Representative data are shown from 1 of 2 independent experiments, with 3 mice per group per experiment. (D) Same as in panel A, except that cells were surface stained for IL-7Rα chain. Representative data are shown from 1 of 3 independent experiments, with 3 mice per group per experiment.