Fig. 6
From: The Wave2 scaffold Hem-1 is required for transition of fetal liver hematopoiesis to bone marrow
c-Abl is essential for engraftment of FL HSCs after homing to marrow. a Schematic of imatinib treatment of 5-week-old adult Hem-1+/+ BM and E14.5 FL Lin− cells. b Imatinib treatment decreased phosphorylation of Crkl in both adult BM and FL Lin− cells (n = 3, **p < 0.01, two-way ANOVA). c and (d) Imatinib treatment decreased cell growth and increased apoptosis only in the Hem-1+/+ E14.5 Lin− FL cells but not adult Hem-1+/+ BM Lin− cells (n = 3, ***p < 0.001, two-way ANOVA). e Schematic of Imatinib treatment of E14.5 Hem-1+/+ Lin− FL cells or 5-week-old adult Hem-1+/+ Lin− BM cells (CD45.2) competitively transplanted into radiation-ablated 5-week-old CD45.1 wild-type mice. The analysis of CD45.2 engraftment in the peripheral blood was performed monthly after transplantation. At 4 months post transplantation, CD45.2 BMCs were assessed to analyze the engraftment. f Imatinib treatment decreased the engraftment Hem-1+/+ E14.5 Lin− FL cells in peripheral blood and bone marrow (n = 5, **p < 0.01, ***p < 0.001, Student’s t test). g Imatinib treatment has no effect on the engraftment ability of adult Hem-1+/+ Lin− BM cells in peripheral blood and bone marrow (n = 5). h Schematic of lentivirally transduced shRNA against c-Abl (c-Abl-shRNA) or vector (CTL shRNA) in Hem-1+/+ E14.5 Lin− CD45.2 FL cells competitively transplanted into radiation-ablated 5-week-old Hem-1+/+ CD45.1 mice. The analysis of CD45.2 engraftment in the peripheral blood was performed monthly after transplantation. At 4 months post transplantation, CD45.2 BMCs were assessed to analyze long-term engraftment. i Lentivirally transduced shRNA against c-Abl reduced engraftment of Hem-1+/+ E14.5 Lin− FL cells into the blood and marrow of recipients (n = 5, ***p < 0.001, Student’s t test). j Schematic of transducing constitutively activated c-Abl (GFP-c-Abl or c-Abl) or vector (GFP-vector or Vector) into Hem-1+/+ and Hem-1−/− E14.5 Lin− cells in a competitive transplantation assay. k Over-expression of c-Abl rescued the defect in the engraftment ability in Hem-1−/− Lin− FL HSCs (n = 5, a p < 0.01 vs. Hem-1+/+ Vector, b p < 0.01 vs. Hem-1−/− GFP-c-Abl, two-way ANOVA)
