Figure 2. Specific silencing of the Matrix GLA protein gene by perfusion of its naked short-interfering RNA to the intact human trabecular meshwork

Post-mortem anterior segments were perfused for three days with either MGP siRNA (OD) or scramble control (OS) at a final concentration of 100 nM. Media with fresh siRNAs was changed on the perfusion syringes every 24 h. After perfusion, trabecular meshwork tissues were dissected and processed for RNA extraction, RT reaction, and real-time TaqMan PCR using MGP and 18S TaqMan probes. Normalization of MGP expression in each sample was achieved using the 18S TaqMan probe hybridized to its own 10^-4 diluted RT. Values of 0 and 100 indicate null and full reduction of the MGP gene expression by MGP siRNA. Scramble negative control samples were normalized to a value of 0 and MGP percent silencing values are expressed as the mean±SEM. MGP expression was significantly and efficiently silenced in the human trabecular meshwork tissues perfused with MGP siRNA as compared to those perfused with scramble control. The asterisk indicates that p=0.00004 for individual 3 (n=6) and that p=0.000002 for individual 4 (n=6).