Validation of the microarray data using Q-PCR
-Fold changes of 13 significantly regulated genes (as determined by the microarray analysis in Table 1) were determined by Q-PCR. Consistent with the microarray data, all -fold changes determined by the Q-PCR analysis were significantly different (p < 0.05) from the control (saline treatment). There was a strong significant correlation between -fold changes obtained by the microarray analysis and those obtained by the Q-PCR analysis (r = 0.979, p < 0.0000001). Results are expressed as mean ± S.D. (n = 3). An additional group of genes (n = 9) that were not significantly (p > 0.05) regulated in the arrays (Table 1) were also tested to confirm the reliability of the statistical approach used for the microarray data analysis; none of these genes was significantly different in the Q-PCR analysis (p > 0.05). For these nine nonsignificantly regulated genes, there was also a strong significant correlation between -fold changes obtained from the microarray analysis and those obtained by the Q-PCR analysis (r = 0.958, p < 0.001) (data not shown).
| Gene Symbol | Gene Title | -Fold Change (Microarray) | -Fold Change (Q-PCR) |
|---|---|---|---|
| Sgk1 | Serum/glucocorticoid regulated kinase | 2.5 ± 0.01 | 2.4 ± 0.1 |
| Gpd1a | Glycerol-3-phosphate dehydrogenase 1 | 2.5 ± 0.02 | |
| 2.1 ± 0.5 | 2.4 ± 0.3 | ||
| Fkbp5a predicted | FK506 binding protein 5 (predicted) | 2.2 ± 0.1 | |
| 2.0 ± 0.03 | 2.2 ± 0.3 | ||
| Abcg2 | ATP-binding cassette, subfamily G (WHITE), member 2 | 2.1 ± 0.1 | 2.1 ± 0.6 |
| Sult1a1 | Sulfotransferase family 1A, phenol-preferring, member 1 | 2.1 ± 0.02 | 1.7 ± 0.1 |
| Mt1a (Metallothionein-1X) | Metallothionein 2610029K21 (predicted) | 2.0 ± 0.01 | 2.0 ± 0.5 |
| Per2 | Period homolog 2 (Drosophila) 2610029K21 (predicted) | 1.8 ± 0.02 | 2.0 ± 0.2 |
| Slc16a1 (MCT1) | Solute carrier family 16 (monocarboxylic acid transporter 1), member 1 | −1.5 ± 0.1 | −1.4 ± 0.2 |
| Tnfrsf11b (Osteoprotegrin) | Tumor necrosis factor receptor superfamily, member 11b | −1.7 ± 0.1 | −1.7 ± 0.02 |
| Adamts1 (ADAM-TS1) | A disintegrin-like and metalloprotease with thrombospondin type 1 motif, 1 | −2.1 ± 0.2 | −1.5 ± 0.1 |
| Tm4sf1 predicted [TM4SF1 (TAAl6)] | Transmembrane 4 superfamily, member 1 (predicted) | −2.1 ± 0.2 | −1.8 ± 0.1 |
| S100a8 (Calgranulin A) | S100 calcium binding protein A8 | −5.5 ± 0.6 | −5.0 ± 0.2 |
| S100a9 (Calgranulin A) | S100 calcium binding protein A9 | −8.2 ± 0.5 | −4.5 ± 0.2 |
↵a Gene represented twice on the microarray chips.