Figure 3. Effects of viral RNA and viral/bacterial DNA on the gene expression of the transcription factor proteins HIF-1α, RELA (p65/NF-κB),
STAT3, and NFAT5. A: Gene expression of transcription factor proteins 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: The mRNA levels were determined with real-time RT–PCR analysis after stimulation of near-confluent (B, D) and confluent (C) cultures for 2, 6, and 24 h (as indicated by the panels of the bars), and are expressed as fold of unstimulated controls.
The cells were stimulated with poly(I:C; 500 µg/ml; B, C) and CpG-ODN (500 nM; D), respectively. Each bar represents data obtained in 3 to 7 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.