Figure 4.
Figure 4. Impact on thymocyte development of targeted deletions of γc or IL-7Rα genes. (A) CD4 versus CD8α profiles of total thymus (left) and gated TCR-βhi thymocytes (right) from Cre− control (black), γc-cKO (red), and IL-7Rα–cKO (blue) mice. FACS profiles for both CD4Cre and E8IIICre-mediated conditional deletion are shown. Numbers indicate frequencies of cells in gates. Data are representative of four independent experiments. (B, Top) Absolute thymocyte numbers for DP, TCR-βhi Int, TCR-βhi CD4SP, and TCR-βhi CD8SP populations are shown for γc-cKO (red) compared with Cre− control mice (black). Cell numbers for both CD4Cre and E8IIICre mice are shown. (B, Bottom) Absolute thymocyte numbers for DP, TCR-βhi Int, TCR-βhi CD4SP, and TCR-βhi CD8SP populations are shown for IL-7Rα–cKO (blue) compared with Cre− control mice (black). Cell numbers for both CD4Cre and E8IIICre mice are shown. Cell numbers were calculated by gating specifically on cells that had deleted γc or IL-7Rα. Data represent the mean of 4–13 individual male and female mice aged 5–9 wk from at least four independent experiments. Error bars represent SEM, and all statistically significant changes are marked with asterisks: **, P < 0.01; ****, P < 0.0001. Any comparisons not marked with asterisks were not significant (P > 0.05). (C, Left) Frequency of MHC-II–selected TCR-βhi CD4SP thymocytes from β2m−/− γc-cKO mice (red) compared with Cre− control mice (black). γc-cKO-derived cells were specifically gated for those that were γc-deleted. Data are depicted as frequencies of total thymus on the left y-axis and relative frequencies normalized to Cre− control on the right y-axis of the histogram plot. Data for E8IIICre mice are shown. (C, Right) Frequency of TCR-βhi CD8SP thymocytes from γc-cKO mice (red), IL-7Rα–cKO mice (blue), and Cre− control mice (black) are shown. γc-cKO and IL-7 Rα-cKO-derived cells were specifically gated for those that were γc- or IL-7Rα–deleted. Data are depicted as frequencies of total thymus on the left y-axis and relative frequencies normalized to Cre− control on the right y-axis of the histogram plot. Data for E8IIICre mice are shown. Means of at least three individual mice are shown, and error bars represent SEM. *, P < 0.05; n.s., not significant. (D) Expression of Qa-2 and HSA on TCR-βhi CD4SP thymocytes from γc-cKO mice (red; specifically gated on γc-deleted cells) and Cre− control mice (black). Shaded histograms represent negative staining control. Data are representative of five independent experiments. (E) Frequency of donor-derived thymocytes from mixed bone marrow chimeras. Mixed bone marrow chimeras were made by reconstituting lethally irradiated mice with a 1:1 mixture of T cell–depleted bone marrow from wild-type and γc-cKO mice. After 8 wk, mice were analyzed for the composition of donor-derived cells within the indicated thymocyte subset using congenic markers to discriminate cells derived from each donor. γc-cKO–derived cells were specifically gated for those that were γc-deleted. The dashed horizontal line indicates the expected frequency of cells derived from each donor. Means of five individual mice are shown, and error bars represent SEM. ***, P = 0.0001.

Impact on thymocyte development of targeted deletions of γc or IL-7Rα genes. (A) CD4 versus CD8α profiles of total thymus (left) and gated TCR-βhi thymocytes (right) from Cre− control (black), γc-cKO (red), and IL-7Rα–cKO (blue) mice. FACS profiles for both CD4Cre and E8IIICre-mediated conditional deletion are shown. Numbers indicate frequencies of cells in gates. Data are representative of four independent experiments. (B, Top) Absolute thymocyte numbers for DP, TCR-βhi Int, TCR-βhi CD4SP, and TCR-βhi CD8SP populations are shown for γc-cKO (red) compared with Cre− control mice (black). Cell numbers for both CD4Cre and E8IIICre mice are shown. (B, Bottom) Absolute thymocyte numbers for DP, TCR-βhi Int, TCR-βhi CD4SP, and TCR-βhi CD8SP populations are shown for IL-7Rα–cKO (blue) compared with Cre− control mice (black). Cell numbers for both CD4Cre and E8IIICre mice are shown. Cell numbers were calculated by gating specifically on cells that had deleted γc or IL-7Rα. Data represent the mean of 4–13 individual male and female mice aged 5–9 wk from at least four independent experiments. Error bars represent SEM, and all statistically significant changes are marked with asterisks: **, P < 0.01; ****, P < 0.0001. Any comparisons not marked with asterisks were not significant (P > 0.05). (C, Left) Frequency of MHC-II–selected TCR-βhi CD4SP thymocytes from β2m−/− γc-cKO mice (red) compared with Cre− control mice (black). γc-cKO-derived cells were specifically gated for those that were γc-deleted. Data are depicted as frequencies of total thymus on the left y-axis and relative frequencies normalized to Cre− control on the right y-axis of the histogram plot. Data for E8IIICre mice are shown. (C, Right) Frequency of TCR-βhi CD8SP thymocytes from γc-cKO mice (red), IL-7Rα–cKO mice (blue), and Cre− control mice (black) are shown. γc-cKO and IL-7 Rα-cKO-derived cells were specifically gated for those that were γc- or IL-7Rα–deleted. Data are depicted as frequencies of total thymus on the left y-axis and relative frequencies normalized to Cre− control on the right y-axis of the histogram plot. Data for E8IIICre mice are shown. Means of at least three individual mice are shown, and error bars represent SEM. *, P < 0.05; n.s., not significant. (D) Expression of Qa-2 and HSA on TCR-βhi CD4SP thymocytes from γc-cKO mice (red; specifically gated on γc-deleted cells) and Cre− control mice (black). Shaded histograms represent negative staining control. Data are representative of five independent experiments. (E) Frequency of donor-derived thymocytes from mixed bone marrow chimeras. Mixed bone marrow chimeras were made by reconstituting lethally irradiated mice with a 1:1 mixture of T cell–depleted bone marrow from wild-type and γc-cKO mice. After 8 wk, mice were analyzed for the composition of donor-derived cells within the indicated thymocyte subset using congenic markers to discriminate cells derived from each donor. γc-cKO–derived cells were specifically gated for those that were γc-deleted. The dashed horizontal line indicates the expected frequency of cells derived from each donor. Means of five individual mice are shown, and error bars represent SEM. ***, P = 0.0001.

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