Figure 7
Figure 7. Role of FLT3 in regulation of multipotent lymphoid-primed progenitors. BM cells from 2- and 19-week-old WT, Fl−/−, and Il7r−/− mice were stained with antibodies against lineage-specific antigens (Lin), KIT, CD34, SCA1, and FLT3. Lymphoid-primed multipotent progenitors (LMPPs) in WT mice were identified as the Lin−SCA1+ KIT+ cells with the highest 25% of FLT3 expression, as previously described.15 (A) Representative FACS profiles of each genotype of 19-week-old mice with gates for LMPPs set based on analysis in WT mice (numbers indicate mean percentages of total LSK cells, from 4-6 mice). (B) Frequencies of LMPPs per total BM cells (2 femora and 2 tibiae) in age-matched 2- and 19-week-old WT, Fl−/−, and Il7r−/− mice. Data represent mean (SD) values of 4 to 6 age-matched mice of each genotype. (C) Quantitative PCR data of indicated genes in FACS-sorted LSKFLT3+ cells from WT and Fl−/− mice. LSKFLT3+ cells were sorted as LSKFLT3hi LMPPs in the case of WT mice as indicated in panel A, whereas for Fl−/− mice the 25% highest FLT3-expressing LSK cells were sorted. Lin−KIT+ BM cells (representing predominantly different lineage-committed progenitors) were used as controls. Data (normalized to the expression of HPRT) are mean (SD) values from 2 independent experiments, each performed in triplicates.

Role of FLT3 in regulation of multipotent lymphoid-primed progenitors. BM cells from 2- and 19-week-old WT, Fl−/−, and Il7r−/− mice were stained with antibodies against lineage-specific antigens (Lin), KIT, CD34, SCA1, and FLT3. Lymphoid-primed multipotent progenitors (LMPPs) in WT mice were identified as the LinSCA1+ KIT+ cells with the highest 25% of FLT3 expression, as previously described.15  (A) Representative FACS profiles of each genotype of 19-week-old mice with gates for LMPPs set based on analysis in WT mice (numbers indicate mean percentages of total LSK cells, from 4-6 mice). (B) Frequencies of LMPPs per total BM cells (2 femora and 2 tibiae) in age-matched 2- and 19-week-old WT, Fl−/−, and Il7r−/− mice. Data represent mean (SD) values of 4 to 6 age-matched mice of each genotype. (C) Quantitative PCR data of indicated genes in FACS-sorted LSKFLT3+ cells from WT and Fl−/− mice. LSKFLT3+ cells were sorted as LSKFLT3hi LMPPs in the case of WT mice as indicated in panel A, whereas for Fl−/− mice the 25% highest FLT3-expressing LSK cells were sorted. LinKIT+ BM cells (representing predominantly different lineage-committed progenitors) were used as controls. Data (normalized to the expression of HPRT) are mean (SD) values from 2 independent experiments, each performed in triplicates.

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