Triptolide-Mediated Apoptosis by Suppression of Focal Adhesion Kinase through Extrinsic and Intrinsic Pathways in Human Melanoma Cells

Figure 3

Effects of TPL on caspases activation in SK-MEL-2 cells. Human SK-MEL-2 cells were treated with 0, 30, 50, and 70 nM of TPL for 24 h, and then cells were collected. (a) Cell lysates were subjected to Western blotting analysis to detect caspase-8, -9, and -3 and PARP. (b and c) Effects of TPL on protein expressions related to apoptosis through Fas death receptor. (b) The cell lysates were subjected to Western blot analysis. Each protein was detected using Fas, FasL, FADD and Bid antibodies. (c) Total RNA was extracted in SK-MEL2 cells treated with TPL. The mRNA levels of Fas receptor and Fas ligand were examined by RT-PCR. (d and e) Effects of TPL on protein expressions related to apoptosis through mitochondria. (d) Each protein was detected using Bcl-2, Bax. (e) The cytosolic and mitochondrial fraction proteins were collected and then detected using cytochrome c antibody. GAPDH and β-actin were used as internal controls.