TABLE 4.

Validation of microarray results by qRT-PCR

Gene name	KyA → RacL11-infected micea			HI-KyA → RacL11-infected miceb
	Fold change by:		P value for qRT-PCR resulte	Fold change by:		P value for qRT-PCR resulte
	Microarrayc	qRT-PCRd		Microarrayc	qRT-PCRd
RSAD2	21.6	34.2	0.0050	5.0	6.7	0.0900
MX1	29.4	43.2	0.0040	5.1	7.9	0.0800
IRF7	27.8	17.9	0.0004	2.2	1.4	0.0610
IFI44L	48.2	45.2	0.0003	2.8	2.3	0.1200
ISG15	19.7	20.4	0.0006	2.6	2.5	0.0920
OAS1a	11.1	12.9	0.0010	4.7	1.5	0.2200
IFI204	29.7	19.9	0.0069	2.5	1.4	0.0348
IFN-γ	8.5	10.6	0.0020	—	0.9	0.3710

^aCBA mice immunized with EHV-1 KyA were infected with pathogenic RacL11 at 3 days postimmunization.

^bCBA mice immunized with HI-KyA were infected with pathogenic RacL11 at 3 days postimmunization.

^cMean fold change from three replicate experiments at 8 h post-RacL11 challenge. —, fold difference of < ±2.0 between infected and mock-infected lungs.

^dThe relative mRNA expression of seven antiviral interferon-stimulated genes (RSAD2, MX1, IRF7, IFI44L, ISG15, OAS1a, and IFI204) and the IFN-γ gene, shown in Table 3, was quantified using qRT-PCR. RT-PCR data were normalized to GAPDH expression.

^eStatistics for the three replicates of each gene were calculated independently. Mean signals were compared by using a t test analysis assuming equal variances and 6 degrees of freedom.