Figure 2.
Sphk2 deletion promotes HSC regeneration after chemotherapy. (A) HSPC (LT-HSC, ST-HSC, MPP, and SLAM HSC) numbers from Sphk2Δ/Δ or control mice at indicated time after 5FU treatment (n = 5 mice per group). (B) Cell cycle and (C) apoptosis analysis of SLAM HSCs from Sphk2Δ/Δ or control mice after 5FU treatment (n = 5 mice per group). (D) Scheme for quantification of functional HSCs via transplantation assay. A total of 2 × 105 BM cells from Sphk2Δ/Δ or control mice on day 7 after 5FU treatment were transplanted into irradiated mice along with 2 × 105 recipient BM cells. (E) PB analysis for total engrafted donor cells at the indicated number of weeks after transplantation and (F) percentage of donor-derived B, T, and myeloid lineage cells 16 weeks after transplantation (WT n = 5-6 mice and Sphk2Δ/Δ n = 4-7 mice). (G) Scheme for quantification of HSC function in response to chemotherapy by transplantation assay. A total of 2 × 105 BM cells from Sphk2Δ/Δ or control mice were transplanted into irradiated mice along with 2 × 105 recipient BM cells. Recipient mice were treated with 5FU at 8 weeks after transplantation. (H) PB analysis for total engrafted donor cells at the indicated number of weeks after transplantation and (I) percentage of donor-derived B, T, and myeloid lineage cells at 28 weeks after transplantation (WT n = 4 to 6 mice and Sphk2Δ/Δ n = 7-10 mice). (J) Survival of Sphk2Δ/Δ, Sphk1Δ/Δ, or control mice after serial 5FU treatment every 7 days (WT n = 10 mice, Sphk1Δ/Δ n = 11 mice, and Sphk2Δ/Δ n = 10 mice). (K) Survival of irradiated recipient mice that received 1 × 106 BM cells from Sphk2Δ/Δ or control mice. Injections (5FU) were performed 8 weeks after transplantation every 7 days (WT n = 10 mice and Sphk2Δ/Δ n = 9 mice). (L) HSPC numbers and (M) cell cycle of HSC from Sphk2Δ/Δ or control mice at day 14 after half-lethal irradiation (n = 5 mice per group). (N) Survival of Sphk2Δ/Δ or control mice after lethal irradiation (n = 10 mice per group). Data represented as mean ± standard deviation. Two-tailed Student t tests were used to assess statistical significance. ∗P < .05, ∗∗P < .01, and ∗∗∗P < .001.

Sphk2 deletion promotes HSC regeneration after chemotherapy. (A) HSPC (LT-HSC, ST-HSC, MPP, and SLAM HSC) numbers from Sphk2Δ/Δ or control mice at indicated time after 5FU treatment (n = 5 mice per group). (B) Cell cycle and (C) apoptosis analysis of SLAM HSCs from Sphk2Δ/Δ or control mice after 5FU treatment (n = 5 mice per group). (D) Scheme for quantification of functional HSCs via transplantation assay. A total of 2 × 105 BM cells from Sphk2Δ/Δ or control mice on day 7 after 5FU treatment were transplanted into irradiated mice along with 2 × 105 recipient BM cells. (E) PB analysis for total engrafted donor cells at the indicated number of weeks after transplantation and (F) percentage of donor-derived B, T, and myeloid lineage cells 16 weeks after transplantation (WT n = 5-6 mice and Sphk2Δ/Δ n = 4-7 mice). (G) Scheme for quantification of HSC function in response to chemotherapy by transplantation assay. A total of 2 × 105 BM cells from Sphk2Δ/Δ or control mice were transplanted into irradiated mice along with 2 × 105 recipient BM cells. Recipient mice were treated with 5FU at 8 weeks after transplantation. (H) PB analysis for total engrafted donor cells at the indicated number of weeks after transplantation and (I) percentage of donor-derived B, T, and myeloid lineage cells at 28 weeks after transplantation (WT n = 4 to 6 mice and Sphk2Δ/Δ n = 7-10 mice). (J) Survival of Sphk2Δ/Δ, Sphk1Δ/Δ, or control mice after serial 5FU treatment every 7 days (WT n = 10 mice, Sphk1Δ/Δ n = 11 mice, and Sphk2Δ/Δ n = 10 mice). (K) Survival of irradiated recipient mice that received 1 × 106 BM cells from Sphk2Δ/Δ or control mice. Injections (5FU) were performed 8 weeks after transplantation every 7 days (WT n = 10 mice and Sphk2Δ/Δ n = 9 mice). (L) HSPC numbers and (M) cell cycle of HSC from Sphk2Δ/Δ or control mice at day 14 after half-lethal irradiation (n = 5 mice per group). (N) Survival of Sphk2Δ/Δ or control mice after lethal irradiation (n = 10 mice per group). Data represented as mean ± standard deviation. Two-tailed Student t tests were used to assess statistical significance. ∗P < .05, ∗∗P < .01, and ∗∗∗P < .001.

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