Figure 6.

Hes1 serves as a putative target of miRNA182-5p in regulating ME%. Human Hes1 has a 7-mer conserved binding site at its 3′-UTR for miRNA182-5p. (a) Flow cytometric analysis of Hes1 expression in anti-miRNA182-5p-transfected K562 cells. The blue and red histograms represent Hes1 expression in scramble control and miRNA182-5p inhibition, respectively. (c–f) Representative images from the colony formation assay in K562 cells (c), miGR1-Hes1- (d), shRNA1-Hes1- (e) and shRNA2-Hes1-nucleofected K562 cells (f), respectively. Images are taken at × 4 magnification. Scale bar represents 100 μm. (g and h) Quantitation of the colony counts from the shRNA- (g) and MiGR1-Hes1-nucleofected K562 cells (h) on day 21. Data are shown as number of colonies per five fields. Error bars show s.e. of three independent experiments with P-values of the mean. *P<0.05, **P<0.01 and ***P<0.001. (i and j) ME% showing percentage of myeloid and erythroid cells in colony formation assay from shRNA- and MiGR1-Hes1-nucleofected K562 cells