Fig. 4

Let-7 miRNAs focused inhibition by TuD demonstrates comparable levels of cell proliferation and equivalent cell morphology compared to control transductions. Cell proliferation was assessed by cell counts (cells/mL) performed at cultures a day 14 and b day 21. Open bars represent control and black bars represent let-7a-TuD. Mean value ± SD of five independent donors for each condition. Cell morphology was analyzed at culture day 21 with the preparation of cytospins followed by Wright–Giemsa staining. Representative images are shown from c control transduction and let-7a-TuD with enucleated cells (inset). d Colony formation assay was performed in control transduction and let-7a-TuD at culture day 14. Cells were transduced with lentivirus particles and cultured in semi-solid methylcellulose medium supplemented with puromycin. Average colony counts were obtained from duplicate wells for each condition from three independent donors. Average percentage of each colony type is shown as separate colors (see color key on the right of the bar graph). C, control transduction; let-7a-TuD, let-7a tough decoy design