Figure 4
Figure 4. Transduction of hematopoietic cells with lentiviral construct contained LEF-1 cDNA led to up-regulation of ELA2/NE. The myeloid cell line U937 was transduced with lentiviral construct with LEF-1 cDNA and green fluorescence protein (GFP) (LEF-1 lv) or only GFP (ctrl lv; A). On day 4 of culture, GFP+ cells were sorted. (B) ELA2 mRNA expression in indicated groups, as measured by qRT-PCR normalized to β-actin, is presented as arbitrary units (AUs); data represent means ± SDs of triplicates (*P < .05, to ctrl lv 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 lv samples). (D) Representative Western blot images showing NE protein expression in total lysates from U937 cells transduced with LEF-1 lv, ctrl lv, or from untransduced U937 cells. Ponceau staining is depicted as a control of the amounts of loaded proteins. (E-F) CD34+ cells from 2 healthy volunteers were transduced with LEF-1 lv GFP or ctrl lv GFP and subsequently incubated without or with G-CSF for 4 days; ELA2 mRNA expression (E) was measured by qRT-PCR normalized to β-actin and is presented as arbitrary units (AUs); data represent means (± SDs) of 2 experiments each measured in triplicates (*P < .05 to G-CSF stimulated ctrl lv 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 < .05 to G-CSF–stimulated ctrl lv samples).

Transduction of hematopoietic cells with lentiviral construct contained LEF-1 cDNA led to up-regulation of ELA2/NE. The myeloid cell line U937 was transduced with lentiviral construct with LEF-1 cDNA and green fluorescence protein (GFP) (LEF-1 lv) or only GFP (ctrl lv; A). On day 4 of culture, GFP+ cells were sorted. (B) ELA2 mRNA expression in indicated groups, as measured by qRT-PCR normalized to β-actin, is presented as arbitrary units (AUs); data represent means ± SDs of triplicates (*P < .05, to ctrl lv 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 lv samples). (D) Representative Western blot images showing NE protein expression in total lysates from U937 cells transduced with LEF-1 lv, ctrl lv, or from untransduced U937 cells. Ponceau staining is depicted as a control of the amounts of loaded proteins. (E-F) CD34+ cells from 2 healthy volunteers were transduced with LEF-1 lv GFP or ctrl lv GFP and subsequently incubated without or with G-CSF for 4 days; ELA2 mRNA expression (E) was measured by qRT-PCR normalized to β-actin and is presented as arbitrary units (AUs); data represent means (± SDs) of 2 experiments each measured in triplicates (*P < .05 to G-CSF stimulated ctrl lv 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 < .05 to G-CSF–stimulated ctrl lv samples).

Close Modal

or Create an Account

Close Modal
Close Modal