Figure 1. NP2-targeting artificial microRNA (amiRNA) down-regulates VEGF-C induced NP2 expression in mouse LECs. A: Double-stranded oligonucleotide encoding a pre-miRNA against mouse NP2. B: Mouse LECs were transfected with NP2-targeting amiRNA (NP2-amiR), negative amiRNA (neg-amiR) or mock-transfected (parental), 48 h after transfection NP2 expression was detected by western blot using anti-NP2 antibody. The images were representative of three independent experiments. β-actin was used as loading control. C: Quantitative real-time PCR analysis of NP2 mRNA in parental, negative amiRNA or NP2 knockdown cells. Data were presented as the mean transcript expression normalized to GAPDH of three independent samples±SEM (n=3 per condition). The asterisk indicates a p<0.01 versus parental control. D: ELISA analysis of NP2 level in mouse corneas at the time point (7 days after injury; n=9 per group). All measurements were performed in duplicate. Bars, SEM. The asterisk indicates a p<0.05 versus saline control.