Effects of Salvianolic Acid B on Protein Expression in Human Umbilical Vein Endothelial Cells
Figure 4
Western blot analysis of caldesmon, phospho-NPM, and vimentin expression after Sal B treatment. (a) HUVECs were pretreated with 125 μg/mL of Sal B for 12 h. The cells were washed and maintained in fresh M199 medium for an additional 12 or 24 h incubation at 37°C. The expression levels of caldesmon, phospho-NPM, and vimentin were assayed. Total tubulin is shown as loading control. These data are representative results from three experiments. The relative amounts of caldesmon and vimentin, compared to total tubulin, and phospho-NPM normalized to the NPM were calculated, and the values represented the mean of triplicate experiments ± standard deviations. Control values were set to 100%. versus control. The bottom panels show that the densities of bands quantified by densitometry (b) HUVECs were pretreated with Sal B at the concentrations of 6.25, 12.5, or 25 μg/mL for 12 h. Cells were then washed and maintained in fresh M199 for an additionally 24 h incubation at 37°C. Dose effects of Sal B on the expression of TM, caldesmon, vimentin, and phospho-NPM protein on HUVECs were demonstrated by Western blot. The tubulin or nucleophosmin was used as normalized control as described above. These data are representative results from three experiments. The bottom panels show the densities of bands analyzed by using the NIH software program ImageJ. versus control.
