Figure 4.
Selective STAT5B activation drives the self-renewal of LSCs. (A) Colony numbers in serial plating assay of HPCLSK cells expressing empty vector (eV), STAT5B, or STAT5BN642H (n = 6/genotype, mean ± SEM). (B) BCR/ABLp210+ LSK retransplantation assay: 2 × 104 BCR/ABLp210-GFP+ FACS-sorted LSK cells were injected IV in NSG mice. Two recipients per donor were injected to use 1 for retransplantation after 13 weeks and 1 for terminal analysis. Experimental workflow (i) and survival analyses of recipients of the primary and secondary transplantation (TP, n = 4/genotype) (ii). (C) pSTAT5 (Y694/699), STAT5A, and STAT5B immunoblot analysis of cytoplasmic (cyt) and nuclear (nuc) fraction of HPC-7 cells expressing JAK2V617F and HPCLSK cells expressing FLT3-ITD or BCR/ABLp210. α-TUBULIN served as loading control for the cytoplasmic fraction and RCC1 for the nuclear fraction. A representative blot of 3 independent experiments is shown. Levels of significance were calculated using 1-way ANOVA (A). **P < .01; ***P < .001.

Selective STAT5B activation drives the self-renewal of LSCs. (A) Colony numbers in serial plating assay of HPCLSK cells expressing empty vector (eV), STAT5B, or STAT5BN642H (n = 6/genotype, mean ± SEM). (B) BCR/ABLp210+ LSK retransplantation assay: 2 × 104 BCR/ABLp210-GFP+ FACS-sorted LSK cells were injected IV in NSG mice. Two recipients per donor were injected to use 1 for retransplantation after 13 weeks and 1 for terminal analysis. Experimental workflow (i) and survival analyses of recipients of the primary and secondary transplantation (TP, n = 4/genotype) (ii). (C) pSTAT5 (Y694/699), STAT5A, and STAT5B immunoblot analysis of cytoplasmic (cyt) and nuclear (nuc) fraction of HPC-7 cells expressing JAK2V617F and HPCLSK cells expressing FLT3-ITD or BCR/ABLp210. α-TUBULIN served as loading control for the cytoplasmic fraction and RCC1 for the nuclear fraction. A representative blot of 3 independent experiments is shown. Levels of significance were calculated using 1-way ANOVA (A). **P < .01; ***P < .001.

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