Impact of DNMT3A variants on proliferation and differentiation of HSPCs. Impact of DNMT3A variants on proliferation and differentiation of HSPCs. (A) Histograms.

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Impact of DNMT3A variants on proliferation and differentiation of HSPCs. Impact of DNMT3A variants on proliferation and differentiation of HSPCs. (A) Histograms of residual CFSE staining to estimate proliferation of CD34+ cells after transfection with shRNAs after 5 d (blue). For comparison, measurements at day of HSPC isolation (day 0, no cell division, shown in red) and unstained controls at day 5 (shown in orange) are provided. (B) KD of transcripts 2 and 4 resulted in higher CFSE retention (slower proliferation) than the control (dashed line; mean ± SD; n = 3). (C) The proportion of CD34 high cells in the fast proliferating fraction was increased upon KD of transcript 2 (gates are indicated in Fig 3A; mean ± SD; n = 3). (D, E) CFU frequency (per 100 initially seeded HSPCs) was analyzed after KD (D) and OE (E) of DNMT3A transcripts (mean ± SD; n = 3). *P < 0.05, **P < 0.01 (t test). Tanja Božić et al. LSA 2018;1:e201800153 © 2018 Božić et al.