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. Author manuscript; available in PMC: 2019 Jun 1.
Published in final edited form as: Stem Cells. 2018 Feb 27;36(6):943–950. doi: 10.1002/stem.2800

Figure 2. HSPCs overexpressing Nfix can withstand cytokine deprivation and display reduced apoptosis during ex vivo culture.

Figure 2

(A) Experimental schematic. 48 hours post-transduction, LSK cells transduced with MND-Control or MND-Nfix were re-plated in 96-well non-tissue culture treated plates in serum-free expansion medium (SFEM) supplemented with either normal (100%) or reduced (25%) levels of STIF cytokines. Every 48-72 hours of culture, cells were counted, assessed for GFP+ cells, and passaged 1:4. Cells were also assessed by flow cytometry for apoptosis via Annexin V and cell cycle via DAPI at day seven of culture. (B) Relative growth of control and NFIX+ cells during ex vivo culture (n = 4). Dotted line indicates the divergence in relative growth between control 100% and control 25% cells. (C) GFP percentage of control and NFIX+ cells during ex vivo culture, assessed by flow cytometry (n = 4). Dotted line indicates significant selection of NFIX+ cells under 25% cytokines. (D) Cell cycle analysis of GFP+ control or NFIX+ cells at day seven of ex vivo culture (n = 3). (E) Percentage of GFP+ apoptotic cells within control or NFIX+ cell cultures at day seven of ex vivo culture (n = 6). All values represent mean ± standard deviation. NS denotes not significant.