Facscan laser

AML cells (2 × 10⁶ cells/ml) were treated with 0.5% DMSO or penfluridol (5 or 7.5 μM) for 24 h. Cells were then fixed in 70% ethanol and stored at − 20 °C overnight, after which they were washed with ice-cold phosphate-buffered saline (PBS) and stained with a propidium iodide (PI) solution (4 μg/m PI, 0.5 mg/ml RNase A, and 1% Triton X-100 in PBS) for 30 min in the dark followed by filtration using a 40-μm nylon mesh (Falcon, San Jose, CA). The DNA contents were measured using a FACScan laser flow cytometer analysis system (Beckman Coulter, Los Angeles, CA). The proportion of nuclei in each phase of the cell cycle was measured, and apoptotic cells were quantified by measuring the sub-G₁ population of the cell cycle.

NSCLC cells were treated with penfluridol for 24 h in the absence and presence of 2DG. The mitochondrial mass of NSCLC cells was determined by selectively loading mitochondria with the red fluorescent dye, MitoTracker Red. Briefly, compound-treated cells were stained with 20 nM MitoTracker Red for 15 min at 37 °C and analyzed by a FACScan laser flow cytometric analysis system (Beckman Coulter, Hialeah, FL, USA).

Cell was cultured and synchronized. After 18 hr of synchronization, serum‐free RPMI medium was replaced by serum‐containing medium, and then, honokiol was treated. After 24 hr treatment, cells were stained with PI (propidium iodide, Sigma Chemical, St. Louis, MO, USA). The PI fluorescence was detected by FACScan laser flow cytometer (Beckman Coulter, Fullerton, CA, USA).