Figure 6: Pten-deficient haematopoietic cells depend on the p110β–Rac1 signalling axis. | Nature Communications

Figure 6: Pten-deficient haematopoietic cells depend on the p110β–Rac1 signalling axis.

From: A PI3K p110β–Rac signalling loop mediates Pten-loss-induced perturbation of haematopoiesis and leukaemogenesis

Figure 6

(a) Rac–GTP activity assay was performed on BM after 7 DPI in the corresponding genotypes. Rac–GTP and total Rac were measured and normalized (n=3; mean with s.e.m.). Two-way ANOVA test was applied to compare the RAC-GTPase/RAC levels. (b) PtenΔ/Δ;p110βΔ/Δ BM cells were transduced with retroviruses expressing HA-tagged wild-type P110β or RBD-mutant P110β, and briefly selected in the presence of puromycin. HA immunoblotting was performed to show expression of RBD-mutant and wild-type P110β (n=3 for each group). (c) Cells from WT control, PtenΔ/Δ or PtenΔ/Δ;p110βΔ/Δ animals were cultured in methylcellulose with myeloid growth factors in the presence of puromycin for 7 days, and colonies were counted at the end of day 7 (n=3 for each group). Two-way ANOVA test was applied to compare the colony numbers. (d) Lineage-negative cells were isolated from WT control, PtenΔ/Δ or PtenΔ/Δ;p110βΔ/Δ mice, and the Rac–GTP activity assay was performed as in (a). Rac–GTP and total Rac were measured and normalized to WT (n=6; mean with s.e.m.). Two-way ANOVA test was applied to compare the RAC-GTPase/RAC levels. **P< 0.01, ***P<0.001

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