Figure 3

Validation of Affymetrix data by quantitative real-time PCR and semi-quantitative PCR analyses. (a) Validation of Affymetrix data by quantitative real-time polymerase chain reaction (PCR) analyses. The fold change was calculated by using the following formula: fold-change = $2^{-(\Delta {\text{C}}_{\text{t}}\text{gene}1-\Delta {\text{C}}_{\text{t}}\text{gene}2)}$. ΔC_t of the gene in the sample in which it is expressed lowest is taken as ΔC_t gene2 to calculate the fold change using the above formula. The resulting fold change is expressed as percentage of the maximum fold change (= 100%) for that particular gene in every assay. Values are expressed as mean ± standard deviation (n = 3; technical replicates). (b) Additional validation of Affymetrix data by semi-quantitative reverse transcription (RT)-PCR analyses. Randomly chosen candidate genes to validate Affymetrix data by semi-quantitiative RT-PCR analyses and their relative expression values expressed as percentage of maximum expression for every gene, as obtained from Affymetrix profiling, are given in the table.