Fig. 3.

miR-335 negatively regulated POU5F1 gene expression. a Bioinformatics study indicated that miR-335 was partially complementary to the 3′-UTR of POU5F1 mRNA. b Different doses of pre-miR-335 were co-transfected with the specific pMIR-REPORT construct into HEK293 cells. Luciferase activities were measured and normalized to the phRL-TK activities. c The effect of pre-miR-335 gradually diminished in 2 weeks. d Osteosarcomas cells were transfected with pre-miR-335, anti-miR-335 or their control respectively. After induction for 7 and 14 days, the cells were harvested for measurement of POU5F1 mRNA using qRT-PCR and e the protein expression using Western blot. Untreated cells was set as control, and β-actin acts as an internal control. All experiments were carried out at least triplicates and the data were presented as the mean ± SD. Student t test was performed to evaluate the difference. *P < 0.05; **P < 0.01