Figure 5. Suppression of vascular endothelial growth factor expression in murine retinas by pSilencer^siVEGF. Murine retinas were extracted and examined for VEGF level. One-way ANOVA followed by the LSD-t-test were used to evaluate significant differences. A p value <0.05 was considered statistically significant. Error bars are standard error of mean (SEM). A: VEGF₁₂₀, VEGF₁₆₄, and VEGF₁₈₈ mRNA levels in murine retinas were examined by RT–PCR. VEGF₁₆₄ mRNA in murine retinas was downregulated after intravitreal administration of pSilencer^siVEGF, whereas VEGF₁₂₀ and VEGF₁₈₈ levels in murine retinas remained unchanged. Lanes are marked as follows: 1) room air-raised mice; 2) murine model with pSilencer^siVEGF injection; 3) murine model with pSilencer null vector injection; and 4) murine model of oxygen-induced retinopathy. B: Relative VEGF isoforms mRNA quantification was related to β-actin mRNA. C: Shown is an immunoblot assay for VEGF₁₆₅ protein in the murine retinas. The total amount of VEGF₁₆₄ immunosignal in murine retinas was reduced by pSilencer^siVEGF. The following abbreviations are used in panel C: ganglion cell layer (GCL), inner nuclear layer (INL), and outer nuclear layer (ONL). D: The levels of VEGF₁₆₅ were quantitated by densitometry and normalized to β-actin protein levels. Data are expressed as mean±SEM and were analyzed by one-way ANOVA followed by LSD-t-test. The means of groups 3 and 4 are statistically greater than the means of groups 1 and 2 (p<0.05).