Time-dependent analysis of the MIA PaCa-2 cells to oxythiamine. A. Two-DE patterns of total proteins from the MIA PaCa-2 cells treated by OT with 0, 12 and 48 h. On the basis of PDQuest software analysis, ratios of normalized spot intensities of four different concentrations of oxythiamine were calculated, and spots showing more than 2.0-fold difference with statistical significance (p < 0.05) were selected after these 2D gels were stained with SYPRO Ruby. B. The temporal profile patterns of 45 differential proteins. The 0 min value was used as a basal time point to calculate the protein fold changes in 12 h and 48 h. After transformed to the logarithmic scale (base 2), the temporal changed profiles of 45 differentially proteins were algorithmically subdivided into nine clusters using K-means clustering method. The protein number of each cluster is indicated. C. The validation of four differential proteins (peroxiredoxin-6, annexin A1, calreticulin and heterogeneous nuclear ribonucleoproteins A2/B1) for the time-dependent group. Western blot scores are annotated under the bands. The results were well consistent with the 2-DE analyses.