Figure 6.

SD70 Inhibits Leukemogenesis In Vitro and In Vivo

(A) Murine leukemia cells were treated with SD70 or DMSO for 3 days with cell viability determined by trypan blue exclusion assay.

(B) Apoptosis analysis on SD70- or DMSO-treated murine leukemia cells.

(C) FACS analysis of myeloid marker Mac1 in murine leukemia cells after SD70 treatment.

(D) Cell-cycle analysis after SD70 treatment.

(E) MLL-AF9-luciferase leukemia cells were transplanted into syngeneic mice and subjected to either vehicle control or SD70 treatment. Bioluminescence imaging was performed at 39 days after transplantation.

(F) Kaplan-Meier survival analysis on the effect of SD70 treatment on MLL-AF9 mediated leukemogenesis (log-rank test p = 0.0143). Median disease latency: control, 55 days; SD70, 61.5 days.

(G) Western blotting analysis of H3K9me3 mark in murine MLL-AF9 leukemia cells after SD70 in vivo treatment. Intensity ratio was determined by densitometry.

(H) Cell viability of human leukemia cell lines were determined 3 days after SD70 or DMSO treatment.

(I) Apoptosis analysis of SD70-treated human leukemia cell lines.

(J) Cell viability of primary AML patient samples after SD70 treatment (MLL, n = 3; non-MLL, n = 4).

(K) NBT reduction assay to determine myeloid differentiation of MLL and non-MLL patient samples. The black bars show the mean.

(L) Primary human MLL leukemia (MLL3) was tagged with luciferase reporter and then transplanted into NSG mice. Bioluminescence imaging of control (n = 6) and SD70-treated (n = 5) cohorts were performed on day 44 after transplantation. Bars show the mean bioluminescence intensity.

(M) Kaplan-Meier survival analysis of vehicle or SD70-treated cohort transplanted with primary human MLL3 leukemia cells (log-rank test p = 0.0018). Median disease latency: control, 59 days; SD70, undefined.

All data shown are mean and SD (n = 3) unless otherwise specified. See also Figure S6.