Journal: British Journal of Cancer

Article Title: α -Tocopheryl succinate promotes selective cell death induced by vitamin K3 in combination with ascorbate

doi: 10.1038/sj.bjc.6605617

Figure Lengend Snippet: Morphological changes of prostate cancer cells exposed to α -tocopheryl succinate ( α -TOS) and vitamin K3 (VK3) plus ascorbic acid (AA) alone or in combination. ( A ) For the soft-agar colony-forming assay, cells were seeded in 24-well culture plates at 10 4 per well in the RPMI-1640 medium containing 0.35% low melting point (LMP) agarose overlaid with 0.7% LMP agarose. The cells were maintained at 37°C in 5% CO 2 for 30 days; every 7 days, 500 μ l of fresh medium was added to each well. The formed colonies were treated with a sub-lethal dose of α -TOS (30 μ M ) or VK3 (3 μ M ) plus AA (0.4 m M ), or the three drugs at these doses together, and the cells were stained with crystal violet after 7 days and inspected by optical microscope. ( B ) PC3 cells were placed overnight in 35-mm dishes on glass coverslips. After a 6-h incubation with α -TOS (30 μ M ) or VK3 (3 μ M ) plus AA (0.4 m M ), or the three drugs at these concentrations together, the cells were stained with phalloidine–TRIC, and the coverslips mounted on microscope slides with Vectrashield containing DAPI and inspected in a fluorescence microscope.

Article Snippet: The cells were then incubated with TRIC-conjugated phalloidine (Sigma) (2 μ g ml −1 ) at room temperature for 30 min, and the coverslips mounted on microscope slides with VectraShield plus DAPI (Vector Laboratories, Burlingame, CA, USA) and inspected in a fluorescence microscope (Zeiss, Axiocam MRc5, Thomwood, NY, USA, magnification × 60).

Techniques: Staining, Microscopy, Incubation, Fluorescence