Figure 5.
Syndecan-2 regulates HSC quiescence and self-renewal capacity. (A) Representative flow cytometric analysis of CD150+CD48– cells within the 34–KSL population treated for 48 hours with two Sdc2 short hairpin RNA clones (shSdc2 Clone #1 and shSdc2 Clone #2) or control short hairpin RNA (shCtrl, n = 3-6 replicates per group). (B) Percentages of CD150+CD48–34–KSL cells (LT-HSCs) in Sdc2– and Sdc2+ cells treated with shSdc2 or shCtrl (n = 3-6 replicates per group). (C) Numbers of phenotypic LT-HSCs after treatment with shSdc2 or shCtrl (n = 3-6 replicates per group). Numbers of total colony-forming cells (CFCs) (D) and CFU-GEMMs (E) in BM 34–KSL cells at 48 hours after treatment with shSdc2 or shCtrl (n = 4 replicates per group). (F) Percentages of total donor CD45.1+ cells in the PB of recipient CD45.2+ mice over time and percentages of donor cells within the B-cell, T-cell, and myeloid cell populations over time after competitive transplantation of shSdc2-treated BM 34–KSL cells or shCtrl-treated BM 34–KSL cells (n = 8-10 recipients per group). (G) Percentages of donor CD45.1+CD150+CD48– cells in the BM at 16 weeks posttransplant in each group (n = 8-10 recipients per group). (H) Representative cell cycle analysis of donor CD45.1+34–KSL cells in the BM of recipient mice at 16 weeks after transplantation of shSdc2-treated KSL cells or shCtrl-treated KSL cells. (I) Quantification of the percentages of donor cells in G0, G1, and G2/S/M phases in the groups shown (n = 8-10 replicates per group). (J) Percentages of total donor CD45.1+ cells, B cells, T cells, and myeloid cells in the PB over time after secondary competitive transplant in the treatment groups shown (n = 8-10 replicates per group). Error bars = standard error of the mean. Statistics denote Holm-Šidák’s post hoc unpaired Student t test after 1- or 2-way analysis of variance; for secondary competitive transplants, statistics denote the overall effect of shSdc2 treatment. *P < .05, **P < .01, ***P < .001, ****P < .0001. ns, not significant; SSC-A, side scatter area.

Syndecan-2 regulates HSC quiescence and self-renewal capacity. (A) Representative flow cytometric analysis of CD150+CD48– cells within the 34–KSL population treated for 48 hours with two Sdc2 short hairpin RNA clones (shSdc2 Clone #1 and shSdc2 Clone #2) or control short hairpin RNA (shCtrl, n = 3-6 replicates per group). (B) Percentages of CD150+CD48–34–KSL cells (LT-HSCs) in Sdc2– and Sdc2+ cells treated with shSdc2 or shCtrl (n = 3-6 replicates per group). (C) Numbers of phenotypic LT-HSCs after treatment with shSdc2 or shCtrl (n = 3-6 replicates per group). Numbers of total colony-forming cells (CFCs) (D) and CFU-GEMMs (E) in BM 34–KSL cells at 48 hours after treatment with shSdc2 or shCtrl (n = 4 replicates per group). (F) Percentages of total donor CD45.1+ cells in the PB of recipient CD45.2+ mice over time and percentages of donor cells within the B-cell, T-cell, and myeloid cell populations over time after competitive transplantation of shSdc2-treated BM 34–KSL cells or shCtrl-treated BM 34–KSL cells (n = 8-10 recipients per group). (G) Percentages of donor CD45.1+CD150+CD48– cells in the BM at 16 weeks posttransplant in each group (n = 8-10 recipients per group). (H) Representative cell cycle analysis of donor CD45.1+34–KSL cells in the BM of recipient mice at 16 weeks after transplantation of shSdc2-treated KSL cells or shCtrl-treated KSL cells. (I) Quantification of the percentages of donor cells in G0, G1, and G2/S/M phases in the groups shown (n = 8-10 replicates per group). (J) Percentages of total donor CD45.1+ cells, B cells, T cells, and myeloid cells in the PB over time after secondary competitive transplant in the treatment groups shown (n = 8-10 replicates per group). Error bars = standard error of the mean. Statistics denote Holm-Šidák’s post hoc unpaired Student t test after 1- or 2-way analysis of variance; for secondary competitive transplants, statistics denote the overall effect of shSdc2 treatment. *P < .05, **P < .01, ***P < .001, ****P < .0001. ns, not significant; SSC-A, side scatter area.

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