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. 2013 Jun 27;3(6):1766–1776. doi: 10.1016/j.celrep.2013.04.031

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© 2013 The Authors

Open Access under CC BY-NC-ND 3.0 license

PMC Copyright notice

Involvement of PU1 Upregulation in the Myeloid Bias of Flt3^ITD/ITD Mice

(A–C) Representative FACS analysis of PU1-YFP expression in lineage-negative (A), LSKCD48⁺150⁻ MPPs (B), and Lin⁻CD19⁻CD24⁻AA4.1⁺CD43^midB220⁺ pre-pro-B (C) cells in BM of 2- to 3-month-old Flt3^+/+ and Flt3^ITD/ITD mice. Also shown is the mean fold difference in PU1-YFP expression for each population from six mice of each genotype.

(D) GSEA demonstrating upregulation of Pu1 target genes in LSKCD48⁺150⁻ MPPs from Flt3^ITD/ITD versus Flt3^+/+ mice (n = 4 of each genotype).

(E) Percentage of B220⁺CD19⁺ in the BM of 6- to 8-week-old Flt3^+/+Pu1^+/+, Flt3^+/+Pu1^+/−, Flt3^ITD/ITDPu1^+/+, and Flt3^ITD/ITDPu1^+/− mice (n = 8–11 mice of each genotype; fold difference and p value are for the difference between Flt3^ITD/ITDPu1^+/+ and Flt3^ITD/ITDPu1^+/− mice).

(F) Percentage of pre-pro-B cells in the BM of 6- to 8-week-old Flt3^+/+Pu1^+/+, Flt3^+/+Pu1^+/−, Flt3^ITD/ITDPu1^+/+ and Flt3^ITD/ITDPu1^+/− mice (n = 3 mice of each genotype).

(G) Percentage of pro-B cells in the BM of 6- to 8-week-old Flt3^+/+Pu1^+/+, Flt3^+/+Pu1^+/−, Flt3^ITD/ITDPu1^+/+ and Flt3^ITD/ITDPu1^+/− mice (n = 3 mice of each genotype).

(H) Percentage of LSKCD48⁺150⁻ MPPs in the BM of 6- to 8-week-old Flt3^+/+Pu1^+/+, Flt3^+/+Pu1^+/−, Flt3^ITD/ITDPu1^+/+ and Flt3^ITD/ITDPu1^+/− mice (n = 7–9 mice of each genotype).

(I) Percentage of Mac1^lowcKit^low myeloid precursor cells in the BM of 6- to 8-week-old Flt3^+/+Pu1^+/+, Flt3^+/+Pu1^+/−, Flt3^ITD/ITDPu1^+/+ and Flt3^ITD/ITDPu1^+/− mice (n = 7–9 mice of each genotype).

(J) GSEA demonstrating upregulation of STAT3 target genes in Flt3^ITD/ITD versus Flt3^+/+ LSKCD48⁺150⁻ MPPs (n = 4 mice of each genotype).

(K and L) Expression of Stat3 target genes in Flt3^+/+ and Flt3^ITD/ITD LSKCD48⁺150⁻ MPPs (K) and pre-pro-B cells (L) (n = 2–4 mice of each genotype, 2–3 replicates per mouse). ND, not detected.

Error bars represent SEM. See also Figure S4.