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. Author manuscript; available in PMC: 2018 Mar 20.
Published in final edited form as: Exp Hematol. 2016 Sep 17;45:74–84. doi: 10.1016/j.exphem.2016.09.003

Figure 1.

Figure 1

Transient knock-down of AKT1 is sufficient to increase human HSPC quiescence. (A) AKT1 expression levels were measured by quantitative real-time polymerase chain reaction analysis 48 hours after siRNA transfection and normalized to GAPDH (***p < 0.001, n = 3). (B) Percentage of human CD34+ cells in expanding medium measured by flow cytometric analysis 7 days after siRNA transfection (p > 0.05, nonsignificant [N.S.], n = 4). (C) Absolute number of human CD34+ cells in expanding medium measured 7 days after siRNA transfection (N.S., n = 4). (D) Flow cytometry analysis of G0 cells using cellular RNA staining (with pyronin Y) and DNA staining (with Hoechst 33342). Percentage of CD34+ cells in the G0 phase of the cell cycle 7 days after siRNA transfection is shown. (E) Quiescence analysis of CD34+ cells 7 days after siRNA transfection (*p < 0.05, n = 4). (F) Frequency of apoptotic CD34+ cells (Annexin V+/7-AAD−) measured 7 days after siRNA transfection (p > 0.05, n = 4). (G) Transient knock-down of AKT1 enhances colony formation of human CD34+ cells. CFC assays were performed using 250 sorted CD34+ cells 7 days after siRNA transfection (***p < 0.001, n = 3).