Fig. (8) Western blot validation of iTRAQ results. Randomly selected
proteins significantly regulated in tianma-stimulated rat aortas
compared with controls. Rats were treated, proteins extracted
and western blot performed as described in materials and methods.
A: Serpinh1, Anxa2 and Gstm2 expressions were up-regulated,
while Vcl and Des expressions were down-regulated. Gapdh, that
did not show any change in the iTRAQ data set, was used as reference
protein. B: Quantitative analyses of the western blots shown in
A. Western blots were performed at least four times for statistical
quantification and analyses (n=4). Values (= relative protein expression)
represent the ratio of densitometric scores for the respective
western blot products and statistical error was indicated as
mean ± SD (*P < 0.05, compared with controls) using the Gapdh
bands as reference. C: The histogram indicates a similar close relationship
between iTRAQ and western blot expression ratios.
Tianma-stimulated and control aorta iTRAQ expression ratios from
selected proteins were consistent with the western blot results and
thus validated a strong agreement in the expression data.