Fig. 3. Effect of the combination treatment of NVP-BKM120 and Prima-1Met on apoptosis of thyroid cancer cells. (a) THJ-29T and FTC-133 cells were treated with BKM120 (1 µM) and Prima-1Met (30 µM). 48 hours after treatment, apoptosis was determined by FACS analysis and presented as percentages of apoptotic cells. (b) Acridine orange/ethidium bromide fluorescence staining was performed in THJ-29T and FTC-133 cells. (c) The levels of cleaved PARP, Bax, Bcl-2 and p21 proteins were analyzed by Western blot. *P<0.05, significant difference between the BKM120/Prima-1Met-treated group and the BKM120 or Prima-1Met-treated group. (d) The release of cytochrome-c from the inter-mitochondrial space into the cytosol was analyzed by immunofluorescence analysis.