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. Author manuscript; available in PMC: 2014 Apr 1.
Published in final edited form as: Aging Cell. 2013 Apr;12(2):303–311. doi: 10.1111/acel.12055

Figure 3. Pro-B cell, but not pre-B cell, growth is inhibited by ABC and TNFα.

Figure 3

Panel A: Isolated pre-B cells (IgM− CD2+ CD19+) and pro-B cells (IgM− CD2− CD19+) from young adult B6 mice were cultured with 5ng/ml rmIL-7 in triplicate IL-7 CFU assays. The IL-7 CFU incidence is provided normalized to 105 input B cell precursors. Panel B: CD2+ pre-B cells (6 × 105) and CD2− pro-B cells (2 × 105) from young adult B6 mice, isolated as in Panel A, were treated with rmTNFα (10ng/ml) in IL-7 CFU assay. Triplicate IL-7 CFU assays were performed. Data are normalized to IL-7 CFU for each cell population, in the absence of TNFα, set to “100”. Panel C: Either old splenic ABC (2 × 105), old bone marrow ABC (2 × 105), or rmTNFα (10ng/ml) were cultured in transwells separated from CD19+ pro-B cells (3 × 105) isolated from RAG-2 gene knockout B6 mice. Culture conditions were as described in Figure 2. IL-7 CFU cultures in each individual experiment were performed in triplicate; each bar represents cumulative results of 2-3 individual experiments with averages and SE shown. In order to compare across experiments, results from each individual experiment were normalized by setting the control (Ctrl) culture IL-7 CFU colony counts (no added ABC or FO B cells or TNFα) to 100. Statistical significance is shown: *, p<0.023 ; **, p< 0.009, ***, p<0.0009.