Lentiviral transduction–induced silencing of ENKUR gene in CD34⁺ stem cells. (A) Flow cytometric analysis of cell-surface markers to confirm the megakaryocytic and platelet fractions after 15 days of culture. (B) RNA expression levels in the CD34⁺ cell–derived MKs and platelets. (C) (i) Western blot of CD34⁺-derived MKs (n = 4 cord blood and n = 4 peripheral blood); (ii) densitometry for the blots. shRNA1 and shRNA2 indicate the 2 shRNA sequences used to silence enkurin (”Materials and Methods”), with a scramble sequence and no silencing as negative and positive controls, respectively. Densitometry was performed using the IVIS imaging software (∗∗∗∗P < .0001; ∗∗∗P < .001; ∗∗P < .01; ∗P < .05, when compared with nonsilenced controls).