Figure 4. Real time PCR validation of array data.

Validation targets were chosen based on significant fold change in the array study at 12 hpi and/or genes previously identified as important in host anti-tick responses [13]. Pre-optimized primers were purchased from Qiagen, and real-time PCR was performed as described in the methods section. All significantly modulated genes at any time point in the validation study are plotted. Significant changes in gene expression at individual time points are marked with an asterisk. Significance was assessed using the delta-delta Ct method for gene expression normalization and measurement, and the linear models in microarray analysis (LIMMA) package for statistical comparisons between infested and tick-free mice, as previously described [13].