Fig. 5

Tg(hsp70:E2A-PBX1-EGFP;lyz:DsRed) transgenic cells with induced acute myeloid leukemia-like disease are transplantable. (A) Immunofluorescent staining of the myeloid-specific marker Lcp1 in frozen sections confirmed myeloid cell invasion of the skeletal musculature in 1-year-old wild-type (WT) (left panel) and Tg(hsp70:E2A-PBX1-EGFP) (right panel) adult fish after 3-month heat shock (yellow arrows show invasion areas of myeloid cells into muscle tissue). (B) Direct observation of engrafted acute myeloid leukemia (AML)-like cells. At 60 days post-transplantation (dpt), AML-like E2A-PBX1 Lyz-DsRed+ cells could be visualized in foxn1/Casper-recipient kidney marrow (KM). The white arrow indicates the engrafted acute myeloid leukemia (AML)-like E2A-PBX1 Lyz-DsRed+ cells. N/N, number of zebrafish larvae showing representative phenotype/total number of zebrafish larvae examined. (C) Wright-Giemsa staining of blood cells in recipient KM after AML-like E2A-PBX1 and WT Lyz-DsRed+ transplantation. The red arrows indicate the AML-like myelocytes, the green arrows indicate the blasts and the black arrows indicate lymphocytes in recipients. (C’) Statistical analysis of cell counts in panel (C). The black asterisks indicate statistical difference (one-way ANOVA, mean ± standard error of the mean; *P<0.05, **P<0.01). (D) Survival curves of Tg(hsp70:E2APBX1-EGFP) and WT fish up to 90 day after heat shock. The black asterisks indicate statistical difference (log-rank [Mantel-Cox] test; **P<0.01). (E) Imaging of dying Tg(hsp70:E2A-PBX1-EGFP;lyz:DsRed) at 15-month-old after 3-month heat shock, reveals severe myeloid cell invasion.