Figure 4. Fold changes in expression of genes not seen to change on the microarray, as shown by qPCR. All qPCR reactions were performed in duplicate on cDNA extracted from 4 separate donors. Genes were amplified using Taqman probes and Taqman Universal PCR master mix (Applied Biosystems). Genes were selected for further qPCR analysis, based on the immune and TGF-β gene groups of interest seen in the microarray analysis. Statistical analysis was undertaken using the REST program (Qiagen), which uses a pair-wise fixed reallocation randomization test to determine significance. Averaged values of duplicate samples from 4 separate donors were statistically analyzed. However, despite appreciable fold-changes in some samples, only TNF and TGF-β2 genes in WNV-infected RPE were statistically different from uninfected RPE. * represents p<0.05.