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. 2020 Oct 5;106(11):2859–2873. doi: 10.3324/haematol.2020.256347

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Copyright© 2021 Ferrata Storti Foundation

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Figure 2. — Exogenous expression of ZNF648 in the induced pluripotent stem cells line HiDEP-1 impedes proliferation and advances differentiation. HiDEP-1 cells were transduced with ZNF648, ZNF648-GFP or a control green fluorescent protein (GFP) construct. (A) Western blot of HiDEP-1 cells probed with ZNF648 antibody (Abcam). Molecular wieght (MW) markers shown on left hand side (B) Images of control and HiDEP-1 cells transduced with ZNF648-GFP; DAPI nuclear staining in blue. (C) Relative fold expansion of ZNF648 and ZNF648-GFP expressing HiDEP-1 cells compared to untransduced (UT) and GFP control cells. *GFP vs. ZNF648, +GFP vs. ZNF648-GFP. **/++ P<0.01; ***/+++ P<0.001, n=2 each for ZNF648 and ZNF648-GFP ± standard deviation, t-test. (D) proportion of HiDEP-1 cells at each stage of differentiation present at day 12 of culture. ProE: proerythroblast; BasoE: basophilic erythroblast; PolyE: polychromatic erythroblast; OrthoE: orthochromatic erythroblast; Retics: reticulocytes. **P<0.01, ***P<0.001, 2-way ANOVA. N=3 ± standard deviation.