Figure 5. Effects of viral RNA and viral/bacterial DNA on the gene expression of inflammatory cytokines. The mRNA levels (B, C, E) were determined with real-time RT–PCR analysis after stimulation of the cells for 2, 6, and 24 h (as indicated by the panels of the bars), and are expressed as fold of unstimulated controls. The level of TNFα protein (D) was determined with ELISA in the cultured media of cells stimulated for 6 and 24 h, and are expressed in percent of unstimulated controls (100%). A: Expression of inflammatory protein genes in cells cultured for 2 (1), 6 (2), and 24 h (3), as determined by RT–PCR. Negative controls (0) were done by adding double-distilled water instead of cDNA as a template. B, D: Near-confluent cultures were stimulated with poly(I:C; 500 µg/ml). C: Dose-dependent effect of poly(I:C). Confluent cultures were stimulated with poly(I:C; 100 and 500 µg/ml, respectively). E: Near-confluent cultures were stimulated with CpG-ODN (500 nM). Each bar represents data obtained in 3 to 5 independent RPE cell lines, each from a different human eye donor; experiments with each cell line were carried out in triplicate. Significant difference versus unstimulated controls: *p<0.05. ●, p<0.05.