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

Figure 1.

Figure 1

Targeting of Prmt1 Suppresses MLL-GAS7 Leukemia

(A) Effect of Prmt1 knockdown on serial replating of transformed cells induced by various leukemic fusions. qRT-PCR analysis of Prmt1 knockdown in leukemic cells. Scale bars represent 0.5 cm.

(B) GST pull-down assays to show the interaction of GAS7 WW domain (WW) with Sam68, PSF, and Prmt1 in vitro.

(C and D) Co-immunoprecipitation of FLAG-MLL-GAS7 with myc-tagged Prmt1, Sam68, and PSF (C) and endogenous Sam68, Prmt1, and PSF (D).

(E) ChIP analysis on the effect of Prmt1 knockdown on H4R3me2as mark and Prmt1 binding in Hoxa9 promoter and gene body region of MLL-GAS7 and E2A-PBX.

(F) qRT-PCR analysis on Hoxa9 and Meis1 expression in MLL-GAS7 after Prmt1 knockdown.

(G) MLL-GAS7 leukemic cells transduced with control or shPrmt1 lentivirus expressing GFP markers. Transduced populations were sorted based on GFP expression and plated into methylcellulose to study colony-forming ability.

(H) Prmt1 knockdown was validated by qRT-PCR, and its effect on H4R3me2as marks was analyzed by western blot with histone H3 as the loading control.

(I) Kaplan-Meier survival analysis of the effect of Prmt1 knockdown on MLL-GAS7 leukemogenesis (log-rank test p < 0.0001). Median disease latency: control, 30 days; shPrmt1, undefined.

(J) Kaplan-Meier survival analysis of mice transplanted with wild type (WT) or Prmt1 knockout (KO) MLL-GAS7 leukemia cells (log-rank test p = 0.0027). Median disease latency: WT, 29 days; Prmt1 KO, undefined.

(K) Western blot analysis of H4R3me2as and H3K4me3 after AMI-408 treatment with histone H3 control for histone loading.

(L) Effect of AMI-408 on colony formation of murine leukemia cell lines.

(M) Kaplan-Meier survival analysis on the effect of AMI-408 treatment on MLL-GAS7 leukemogenesis (log-rank test p = 0.0341). Median disease latency: control, 21 days; AMI-408, 27.5 days.

All data shown are mean and SD (n = 3) unless otherwise specified. See also Figure S1. For all figures, asterisks indicate ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001; ns, not significant.