Effects of mimic-miR-720 on stem cell phenotype of DPCs.

A) Quantitative assessment of miR-720 levels. *** P<0.001, One way ANOVA/Tukey. Results are representative of at least 3 independent experiments. B) mimic-miR-720 reduced mRNA levels of NANOG, but increased the levels of DNMT3B and DNMT1 mRNA. No significant changes were observed in DNMT3A mRNA upon miR-720 transfection. * P<0.05, ** P<0.01, NS =  non-significant, unpaired t-test, compared to si-control group. Results are representative of at least 3 independent experiments. C) Representative images of immunocytochemistry for NANOG protein levels. D) Quantitative analysis of NANOG or DNMTs positive cells. Mimic-miR-720 significantly reduced the number of NANOG positive cells, while increasing those for DNMT3B. There was no significant change in the percentages of DNMT1 or DNMT3A positive cells. Results in C–D are representative of at least 3 independent experiments. Quantitative analysis was performed on 500 cells/well, with at least triplicate samples. * P<0.05, ** P<0.01, NS =  non-significant, unpaired t-test, compared to si-control group. G) Transfection of mimic-miR-720 decreased the percentage of SSEA-4⁺ cells. Results are representative of 3 independent experiments.