Fig. 3.

Validation of iTRAQ-detected quantitative differences in protein levels in trisomy 21. A, confirmation of the differential expression of CSE1L, SERPINH1, ANXA5, and TAGLN by Western blotting. Total protein lysates were used from three independently grown 47-1 and D3 cell lines. B, Western blotting of WA17 (mouse fibroblast line with supernumerary HSA21) and A9 (its euploid control) lysates. Antibodies against CSE1L and SERPINH1 were used. C, the expression data (bar chart) contains information from quantitative RT-PCR from n = 7 47-1/D3 pairs (four pairs were the same samples as were applied on the Affymetrix MG-U74Av2 mouse array experiments, and three additional RNA samples were from the same 47-1/D3 pairs as in the Western analysis). Error bars are indicated. ** shows p < 0.01 (highly significant).