Figure 4.
DEK overexpression impairs HSC maintenance and self-renewal. (A) Representative flow plots of HPCs, LSK, CD34−LSK, and HSCs (CD34−CD135−LSK or CD48−CD150+LSK) analysis in BM from DekCon and DekTg mice. (B) The bar graphs show the cell count of indicated cells in BM (n = 5–9 mice per group). (C) Experimental schematic for serial transplantation assay. A total of 1 × 106 BM cells from CD45.2 littermates were transplanted into lethally irradiated recipient mice (CD45.1); after 16 wk, 1 × 106 chimeric BM cells from the primary recipients were transplanted into the second recipient mice. (D) Survival curve of recipient mice for the 1-first (upper) and 2-second (bottom) transplantation (n = 8 recipient mice per group). (E) Representative flow plots of HPCs, LSK, and HSCs (CD48−CD150+LSK) analysis in BM from the 1-first recipient mice. (F) The bar graphs show the cell count of HPCs, LSK, and HSCs in BM from the 1-first recipient mice (n = 4–6 recipient mice per group). (G) Experimental schematic for the competitive transplantation assay. A total of 1 × 106 BM cells from DekTg-fl or DekTg-flMx1-Cre mice (donor CD45.2) were transplanted into lethally irradiated recipient mice (CD45.1/CD45.2), together with competitor BM cells (1 × 106) from WT mice (CD45.1). 4 wk after transplantation, the recipient mice were injected with pIpC (10 mg/kg, every other day, thrice) to induce DEK overexpression, followed by injection with 100 mg/kg HU or saline daily for 4 wk. (H) Representative flow plot of chimerism analysis in PB from recipient mice. The chart on the right shows the donor chimerism in PB of recipient mice at indicated time points after transplantation (n = 6 recipient mice per group). (I) The bar graphs show the donor chimerism in BM (left), LSK (middle), and HSCs (right) of recipient mice at 20 wk after transplantation (n = 6 recipient mice per group). The data presented in panels A–I are representative of two independent experiments. Data are shown as mean ± SD. Statistical analyses were performed using a two-tailed unpaired Student’s t test for panels B and F, a Cox–Mantel test for panel D, and one-way ANOVA followed by Tukey’s post hoc test for panels H and I. P values are presented as follows: *P < 0.05, ***P < 0.001. Refer to the image caption for details.

DEK overexpression impairs HSC maintenance and self-renewal. (A) Representative flow plots of HPCs, LSK, CD34−LSK, and HSCs (CD34−CD135−LSK or CD48−CD150+LSK) analysis in BM from DekCon and DekTg mice. (B) The bar graphs show the cell count of indicated cells in BM (n = 5–9 mice per group). (C) Experimental schematic for serial transplantation assay. A total of 1 × 106 BM cells from CD45.2 littermates were transplanted into lethally irradiated recipient mice (CD45.1); after 16 wk, 1 × 106 chimeric BM cells from the primary recipients were transplanted into the second recipient mice. (D) Survival curve of recipient mice for the 1-first (upper) and 2-second (bottom) transplantation (n = 8 recipient mice per group). (E) Representative flow plots of HPCs, LSK, and HSCs (CD48−CD150+LSK) analysis in BM from the 1-first recipient mice. (F) The bar graphs show the cell count of HPCs, LSK, and HSCs in BM from the 1-first recipient mice (n = 4–6 recipient mice per group). (G) Experimental schematic for the competitive transplantation assay. A total of 1 × 106 BM cells from DekTg-fl or DekTg-flMx1-Cre mice (donor CD45.2) were transplanted into lethally irradiated recipient mice (CD45.1/CD45.2), together with competitor BM cells (1 × 106) from WT mice (CD45.1). 4 wk after transplantation, the recipient mice were injected with pIpC (10 mg/kg, every other day, thrice) to induce DEK overexpression, followed by injection with 100 mg/kg HU or saline daily for 4 wk. (H) Representative flow plot of chimerism analysis in PB from recipient mice. The chart on the right shows the donor chimerism in PB of recipient mice at indicated time points after transplantation (n = 6 recipient mice per group). (I) The bar graphs show the donor chimerism in BM (left), LSK (middle), and HSCs (right) of recipient mice at 20 wk after transplantation (n = 6 recipient mice per group). The data presented in panels A–I are representative of two independent experiments. Data are shown as mean ± SD. Statistical analyses were performed using a two-tailed unpaired Student’s t test for panels B and F, a Cox–Mantel test for panel D, and one-way ANOVA followed by Tukey’s post hoc test for panels H and I. P values are presented as follows: *P < 0.05, ***P < 0.001.

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