Transduction of hematopoietic cells with LEF-1 shRNA resulted in down-regulation of ELA2/NE. The myeloid cell line U937 was transduced with lentiviral constructs with red fluorescence protein (RFP) and 2 different LEF-1–specific shRNAs (LEF-1 shRNA 1; LEF-1 shRNA 2) or with irrelevant shRNA (ctrl shRNA; A). On day 4 of culture, we sorted and analyzed RFP⁺ cells. (B) ELA2 mRNA expression in indicated groups, as measured by qRT-PCR normalized to β-actin and is presented as arbitrary units (AUs); data represent means ± SDs of triplicates (*P < .05 to ctrl shRNA samples). (C) NE protein secretion into culture supernatants was assessed with the use of NE-specific ELISA. Data represent means ± SDs and are derived from 2 independent experiments each measured in triplicate (*P < .05 to ctrl shRNA samples). (D) Representative Western blot images from the same gel and same experiment showing NE protein expression in total lysates from U937 cells transduced with shRNA constructs, as indicated above. β-Actin staining was used as a control of the amounts of loaded proteins. Vertical lines have been inserted to indicate a repositioned gel lane. (E-F) CD34⁺ cells from 2 healthy volunteers were transduced with lentiviral constructs with RFP and LEF-1–specific shRNA (LEF-1 shRNA 1) or with irrelevant shRNA (ctrl shRNA); ELA2 mRNA expression (E) was measured by qRT-PCR normalized to β-actin and is presented as AUs. Data represent means ± SDs of 2 experiments measured in triplicates (*P < .05 to G-CSF–stimulated ctrl shRNA samples). NE protein secretion into culture supernatants (F) was assessed with the use of NE-specific ELISA; data represent means ± SDs and are derived from 2 independent experiments each measured in triplicate (**P < .01 to G-CSF–stimulated ctrl shRNA samples).