Figure 1
Gene-expression profiles of HSC polarity fate determinants. Candidate genes were classified according to Gene Ontology (GO) groups (see below) based initially on biologic process (Groups I-XI). In the absence of a biologic process, they were then based on molecular function (Group XII) and cellular component (Group XIII). The expression of these candidate genes was performed on purified HSC populations compared with more differentiated progenitors. Each dataset is represented by the mean average δCt from 2 experiments in which for each respective reference range, the lowest numbers (green bars) correspond to the highest transcript copy numbers. Shown are gene expressions from (A) HSCs sorted on CD150+48−41−Lin− (with a stem cell frequency of 1 in 10 based on CRUs; data not shown) and progenitors CD150+48+41−Lin− (B) HSCs sorted on CD49b−rhodaminelow LSK (with a stem cell frequency of 1 in 8 based on CRUs, data not shown) and progenitors CD49b−rhodaminehigh LSKs, and (C) 2 independent murine leukemias, FLA2 and FLB1, derived from HoxA9+Meis1 transduced FL cells, that differ in leukemia stem cell frequency of, respectively, 1 in 1.4 and 1 in 347 (based on previous CRU, data not shown). (D) A subset of these candidate genes were profiled after CD150+48−41−Lin− HSCs were transduced with the established expansion factor HoxB4 and compared with MSCV vector alone transduced HSCs. GO Groups: I_GO_7049 cell cycle; II_51301 cell division; III_30154 cell differentiation; IV_45165 cell-fate commitment; V_7163 establishment or maintenance of cell polarity; VI_16192 vesicle mediated transport; VII_16459 myosin complex; _7017 microtubule-based process; VIII_7154 cell communication; IX_50678, _7242, _7169, _6468 receptor signaling pathways; X_48729 tissue morphogenesis; and, _43066 negative regulation of apoptosis; XI_33036, _51234 macromolecule and establishment of localization; XII_5515, _8092 protein and cytoskeleton binding; _3774 motor activity; _8266, _3723 RNA binding; and XIII_5886,16 020 membrane; _45177 apical part of cell. From studies shown in panels A through C, a scoring criteria (data not shown) based on the absolute average δCt and the relative expression in the stem versus progenitor cell populations was used to quantify the candidates chosen for the in vitro to in vivo overexpression screen (see also supplemental Table 1).

Gene-expression profiles of HSC polarity fate determinants. Candidate genes were classified according to Gene Ontology (GO) groups (see below) based initially on biologic process (Groups I-XI). In the absence of a biologic process, they were then based on molecular function (Group XII) and cellular component (Group XIII). The expression of these candidate genes was performed on purified HSC populations compared with more differentiated progenitors. Each dataset is represented by the mean average δCt from 2 experiments in which for each respective reference range, the lowest numbers (green bars) correspond to the highest transcript copy numbers. Shown are gene expressions from (A) HSCs sorted on CD150+48−41−Lin− (with a stem cell frequency of 1 in 10 based on CRUs; data not shown) and progenitors CD150+48+41−Lin− (B) HSCs sorted on CD49b−rhodaminelow LSK (with a stem cell frequency of 1 in 8 based on CRUs, data not shown) and progenitors CD49b−rhodaminehigh LSKs, and (C) 2 independent murine leukemias, FLA2 and FLB1, derived from HoxA9+Meis1 transduced FL cells, that differ in leukemia stem cell frequency of, respectively, 1 in 1.4 and 1 in 347 (based on previous CRU, data not shown). (D) A subset of these candidate genes were profiled after CD150+48−41−Lin− HSCs were transduced with the established expansion factor HoxB4 and compared with MSCV vector alone transduced HSCs. GO Groups: I_GO_7049 cell cycle; II_51301 cell division; III_30154 cell differentiation; IV_45165 cell-fate commitment; V_7163 establishment or maintenance of cell polarity; VI_16192 vesicle mediated transport; VII_16459 myosin complex; _7017 microtubule-based process; VIII_7154 cell communication; IX_50678, _7242, _7169, _6468 receptor signaling pathways; X_48729 tissue morphogenesis; and, _43066 negative regulation of apoptosis; XI_33036, _51234 macromolecule and establishment of localization; XII_5515, _8092 protein and cytoskeleton binding; _3774 motor activity; _8266, _3723 RNA binding; and XIII_5886,16 020 membrane; _45177 apical part of cell. From studies shown in panels A through C, a scoring criteria (data not shown) based on the absolute average δCt and the relative expression in the stem versus progenitor cell populations was used to quantify the candidates chosen for the in vitro to in vivo overexpression screen (see also supplemental Table 1).

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