Validation of protein targets identified by 2D DIGE. Ex vivo monocytes were treated with 200 µg/mL of individual IgG samples: 10 VT, 8 PM, or 10 HC for 6 hours and levels of mRNA measured by qPCR (A-D). Data points represent the fold change expression of each sample compared with untreated; mean of duplicates and standard errors are displayed. Data are representative of at least 3 independent experiments. Representative western blots of monocytes treated with the 10 VT, 8 PM, and 10 HC IgG samples are shown (E-F). Blots were scanned and analyzed densitometrically. Graphical representations of the density ratios of each protein and GAPDH expressed in arbitrary scanning units are displayed (G-H). Data are representative of at least 2 independent biological replicates. Statistically significant difference (*P < .05, **P < .005) was determined by ANOVA. 2D, 2-dimensional; ANOVA, analysis of variance; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; qPCR, quantitative PCR.