Figure 4: p110β deletion rescued long-term HSC (LT-HSC) reconstitution and suppressed leukaemia initiation and homing defects after Pten deletion.
(a) Competitive repopulation experiment: bone marrow (BM) cells were isolated at 7 days post pIpC (DPI) from CD45.2+ ctrl, PtenΔ/Δ, PtenΔ/Δ;p110αΔ/Δ and PtenΔ/Δ;p110βΔ/Δ animals, and transplanted into lethally irradiated recipient animals along with CD45.1+ BM cells. The donor chimaerism in the peripheral blood was measured at regular intervals for 20 weeks, represented as the percentage of CD45.2+ cells among all leukocytes in the peripheral blood (n=9 for all groups). Two-way ANOVA test was applied to compare the donor derived cell numbers. (b) Disease-free survival of animals in the competitive repopulation experiment. Statistical analysis was performed at week 20 post transplantation using Fisher’s exact test. All samples were compared with the control group. Fischer exact test was applied to test overall survival. (c) After 20 weeks, the competitive repopulation experiment was terminated, and the remaining numbers of LSK, ST-HSC and LT-HSCs in the BM were measured by flow cytometry (n=4 for each group except n=5 for PtenΔ/Δ;p110δ−/−). Two-way ANOVA test was applied to compare the donor derived stem cell numbers. (d) Homing potential of CD45.2-positive carboxyfluorescein diacetate succinimidyl ester (CFSE)-labelled control, PtenΔ/Δ and PtenΔ/Δ;p110βΔ/Δ cells in 24 h in irradiated recipients (n=5) for each group. Two-way ANOVA test was applied to compare the donor derived stem cell numbers. *P<0.05, **P< 0.01, ***P<0.001
