Proteomic Approach to Evaluate Mechanisms That Contribute to Food Allergenicity: Comparative 2D-DIGE Analysis of Radioallergosorbent Test Positive and Negative Patients
Figure 5
Relative gene expressions of IL-4, IL-17, IL-10, and MCEM1 in RAST+ versus RAST− patients as quantitated by QPCR. Gene expression analysis of IL-4, IL-17, IL-10, and MCEM1 was done in RNA extracted from PBMC from RAST+ and RAST− subjects. RNA was reverse transcribed and mRNA expression levels of the above genes were quantitated using real time QPCR. Relative expression of mRNA species was calculated using the comparative CT or the ΔΔCt method. This method calculates fold changes between each normal sample and its paired affected sample. ΔCt represents normalized gene expression level (normalized to its own internal control housekeeping gene β-actin). ΔCt for test and control samples were averaged and compared to obtain the ΔΔCt value (an average value). CV, or coefficient of variation, is calculated using the formula STDEV/average. Data are the mean ± SD of 3 separate experiments done in duplicate. Data are presented as TAI or the transcript accumulation index. Statistical significance was determined by Student’s -test based on comparison between the RAST+ versus RAST− patients.
