Skip to main content
. 2016 Jan 11;29(1):32–48. doi: 10.1016/j.ccell.2015.12.007

Figure 2.

Figure 2

Recruitment of Prmt1 by MOZ-TIF2 Is Necessary but Not Sufficient for HSPC Transformation

(A) Co-immunoprecipitation of FLAG-MOZ-TIF2 (fMT2) with myc-Prmt1 (upper) and endogenous Prmt1 (lower).

(B) ChIP analysis of H4R3me2as and Prmt1 localization on Hoxa9 promoter and gene body in MOZ-TIF2 after Prmt1 knockdown (KD).

(C) Co-immunoprecipitation of different FLAG-MOZ-TIF2 deletion mutants (indicated in D) and myc-Prmt1.

(D) RTTA to study the effect of N-terminal deletion mutants, active and inactive Prmt1 (P1) rescue fusions of MOZ-TIF2 on leukemic transformation.

(E) ChIP analysis of H4R3me2as mark on HOXA9 loci in HEK293 cells transfected with WT MOZ-TIF2 or ΔN79.

(F) Kaplan-Meier survival analysis of the effect of Prmt1 knockdown on MOZ-TIF2-mediated leukemogenesis (log-rank test p ≤ 0.0001). Bioluminescence imaging was performed at 21 days after transplantation. Median disease latency: control, 33 days; shPrmt1, undefined.

(G) Kaplan-Meier survival analysis of mice transplanted with WT or Prmt1 KO MOZ-TIF2 leukemia cells (log-rank test p = 0.0031). Median disease latency: WT, 35 days; Prmt1 KO, undefined.

(H) Western blot analysis on the effect of AMI-408 on H4R3me2as, H3K4me3, and the loading control histone H3. Band intensity ratio was determined by densitometry and normalized to vehicle control.

(I) Bioluminescence imaging of mice transplanted with MOZ-TIF2-luciferase leukemic cells 3 weeks after AMI-408 or carrier treatment.

(J) Kaplan-Meier survival analysis of AMI-408 and control treatment on MOZ-TIF2 leukemogenesis (log-rank test p = 0.0042). Median disease latency: control, 35.5 days; AMI-408, 48 days.

(K) Morphology of third-round colony of HSPC transformed by MOZ-TIF2 and its Prmt1 rescue fusion. FACS analysis of the transformed cells stained with c-kit, Gr1, and Mac1. Scale bars represent 50 μm.

(L) Kaplan-Meier survival analysis of mice transplanted with MOZ-TIF2 or MT2ΔN79ΔAD2-P1 transformed cells (log-rank test p < 0.0001). Median disease latency: MOZ-TIF2, 69 days; MT2ΔN79ΔAD2-P1, undefined.

All data shown are mean and SD (n = 3) unless otherwise specified. See also Figure S2.