Figure 2

Suppression of VEGF-A and ICAM-1 expression by CODEMIR-1 and -2. Unless otherwise indicated, VEGF-A (ELISA) or ICAM-1 (FACS) were assayed in ARPE-19 cells 48 hours post-transfection and 24 hours post-stimulation with 130 μM Deferoxamine or 1 ng/mL IL-1β respectively. All data points indicate the mean of triplicate samples. Error bars show standard deviation. Statistical significance was determined by two-way ANOVA using a Bonferroni post-test (* p < 0.001 as compared to either untransfected and/or irrelevant siRNA; ** p < 0.001 as compared to untransfected and p < 0.01 as compared to irrelevant siRNA; † p < 0.001 compared to CODEMIR-1). (a) Gene suppression by CODEMIR-1 and -2. (b) Dose responsiveness of gene suppression by CODEMIR-1. Cells were transfected at the indicated concentrations. (ND = not determined). (c) Effect of mismatches on gene suppression by CODEMIR-1. (d) Suppression of VEGF-A and ICAM-1 reporter constructs by CODEMIR-1ARPE-19 cells were co-transfected with the AmCyan reporter and AsRed control plasmids and 40 nM indicated RNA duplexes. Fluorescence was assessed by FACS 48 hours post-transfection. (e) Suppression of VEGF-A and ICAM-1 mRNA expression by selected CODEMIRs. For VEGF-A, stimulation was performed 24 hours post-transfection using 65 μM Deferoxamine and the QuantiGene^® assay was performed on cell lysates prepared 24 hours post-stimulation. For ICAM-1 the QuantiGene^® assay was performed on cell lysates prepared 24 hours post-transfection.