Fig. 3

TSA treatment induces the intrinsic apoptotic pathway in LECs. a Cells were exposed to 400 nM TSA for 24 h as indicated. For apoptosis determination, we used a colorimetric assay that quantified histone complexed DNA fragments. Average absorbance values (mean ± SE) from 3 wells per experimental condition are displayed; data are expressed as apoptotic cells in percentage (%) with regard to solvent controls (=100%; ethanol). *p < 0.05 vs Ctrl. b and (c) Representative western blot analyses of LECs that were left untreated (solvent only) or were treated with TSA in a concentration- and time-dependent manner. Apoptosis related proteins, like cleaved caspases 9, 3 and 7, cIAP-1/2 and α-Tubulin as loading control were detected by enhanced chemiluminescence. Comparable results were obtained from three independent experiments. d LECs were left untreated (solvent only, Ethanol) or were treated with 400 nM TSA for 24 h. The release of cyctochrome c to cytosol was measured using the cytochrome c ELISA. Mean values from four independent experiments are depicted ± SE (error bars). *p < 0.05 vs Ctrl