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. 2015 Feb 9;212(2):203–216. doi: 10.1084/jem.20132544

Figure 7.

Figure 7.

Rescue of the HSC phenotype in A20Hem-KO IFN-γ−/− mice. (A) Representative images of control, IFN-γ−/−, A20Hem-KO, and A20Hem-KO IFN-γ−/− (DKO) mice showing body size (n = 5). (B) Bar graphs showing body size of control (n = 9), A20Hem-KO (n = 3), and DKO (n = 4) mice. (C) Kaplan-Meier survival curve analysis of control (n = 26), IFN-γ−/− (n = 11), A20Hem-KO (n = 10), and DKO (n = 15) mice. Significance (****, P < 0.0001) was assessed using the log-rank test. (D) FACS plots showing immunophenotype of LSK, LT-HSCs, ST-HSCs, and MPPs of 14-d-old control, IFN-γ−/−, A20Hem-KO, and DKO mice. Data are representative of two independent experiments. (E) Absolute numbers of LT-HSCs in the BM (two femurs and two tibias) of 14-d-old animals. Data are pooled from two independent experiments with two mice per group. (F) Frequencies of Sca1-expressing cells in the Lin−c-Kit+CD150+ fraction of the BM of 14-d-old animals. Data are pooled from two independent experiments with two mice per group. (G) Kaplan-Meier survival curve analysis of WT recipients after BMT. Data are pooled from two independent experiments with 11 mice each for control, A20Hem-KO, and DKO groups and 6 mice for the IFN-γ−/− group. Significance (*, P < 0.03) was assessed using the log-rank test. (H) Frequencies of donor (CD45.2+) cells in the peripheral blood of WT congenic recipients at 5 wk after transplantation. Data are representative of two independent experiments and are pooled from six mice each for control and IFN-γ−/− groups, five mice for the A20Hem-KO group, and seven mice for the DKO group. (I) Frequencies of donor (CD45.2+) cells in the peripheral blood of WT congenic recipients at 14 wk after transplantation. Data are representative of two independent experiments and are pooled from five mice for control and four mice for DKO groups. (J and K) Frequencies of donor (CD45.2+) hematopoiesis in the peripheral blood of WT recipients, at 4 wk after transplantation. Recipients received mixed chimera containing donor (CD45.2+) BM cells (from A20Hem-KO, DKO, and control mice) and WT competitor (CD45.1+) BM cells at a ratio of 1:1 (J) and 5:1 (K), respectively. Data are representative of two independent experiments and are pooled from five mice each for A20Hem-KO and DKO groups and four mice for control group (J) and five mice for DKO group, three mice for A20Hem-KO group, and five mice for control group (K). (L and M) Frequencies of donor-derived cells in total CD11b+ (L) and CD19+ (M) fractions in the peripheral blood of WT recipients from the 1:1 group of J. Data are representative of two independent experiments and are pooled from five mice each for A20Hem-KO and DKO groups and four mice for the control group. All data represent mean ± SEM. Two-tailed Student’s t tests were used to assess statistical significance (*, P < 0.05; **, P < 0.01; ***, P < 0.001).