Figure 5
AE9a but not AE9aNT causes myeloid leukemia and a typical leukemic differentiation pattern. Upon 5-FU injection, BM progenitor cells, transduced with either IRES-GFP (control), AE9a-IRES-GFP (AE9a) or AE9aNT-IRES-GFP (AE9aNT), were transplanted into lethally irradiated (11Gy) C57BL/6 Ly5.1 mice (see overview in Supplementary Figure S4a). (a, left) Kaplan–Meier plot showing that the majority of AE9a transplanted mice died between day 150 and 200 post transplantation whereas AE9aNT mice and control animals did not succumb to leukemia. (a, right) Representative images of BM cells stained with Giemsa from AE9a or AE9aNT mice analyzed 214 days post-transplantation. While AE9a mice presented with homogenous population of immature blasts, AE9aNT BM cells contained heterogenous and differentiated populations. (b) c-kit expression in GFP+ cells in PB was monitored at 6–8 weeks intervals via flow cytometry. AE9a transplanted mice displayed significantly elevated c-kit+ cell numbers compared to GFP or AE9aNT transplanted mice over the entire observation period. (nsNot significant, *P<0.05, **P<0.01, unpaired Student’s t-test). (c) Analysis of spleen and BM cells 214 days post-transplantation. (c, left) WBC counts from the spleen of mice transplanted with AE9a (red) are significantly elevated compared to mice transplanted with GFP (control, green) and AE9aNT (blue) (nsNot significant, **P<0.01, unpaired Student’s t-test, k=103). Mean values based on the number of animals. (c, right) Flow cytometric analysis of c-kit expression in BM and spleen of representative GFP (control, green), AE9a (red) and AE9aNT (blue) transplanted mice indicate a higher percentage of c-kit+ cells in BM and spleen of AE9a transplanted mice.
