Research Article

Explore the Molecular Mechanism of Apoptosis Induced by Tanshinone IIA on Activated Rat Hepatic Stellate Cells

Figure 3

(a) Typical two-dimensional gel electrophoresis (2-DE) pattern of HSC-T6 cells treated with 7.5 μM Tan IIA for 24 h. The protein lysate (300 μg) was focused on a pH 4–7 linear IPG strip before being separated on a 10% polyacrylamide gel. Protein spots with significant changes in intensity are labeled with Arabic numerals. (b) Three-dimensional images of protein spots with up- or downregulated expression. The blue lines delineate the edge of each protein. (c) An MALDI-TOF spectrum of trypsinized prohibitin and the parent ions m/z 1149.5934, 1396.8345, and 1460.6582 were selected for further analysis by an Ultraflex MS/MS operated in the LIFT mode using FlexControl software. The amino acid sequences were unambiguously assigned to the rat prohibitin protein. A sequence was confirmed from the labeled b- and y-ions in the spectrum. (d) Western blot analysis was applied to validate protein changes revealed by 2-DE analysis. GAPDH was used as an internal control, and the relative expression to GAPDH was shown at the bottom. (e) Semiquantitative RT-PCR. Tan IIA treatment upregulated the mRNA expression of prohibitin. -actin was used as an internal control. Each bar represents the mean ± SD calculated from 3 independent experiments. * compared to the control. (f) Biologic network analyses of differentially expressed proteins using MetaCore mapping tools. Top 10 pathways are ranked based upon -value, and bars represent inverse log of the -value.