The cells were relatively resistant to TRAIL-induced cell death; the viability curve reached a plateau at 0.5 μg ml−1, and, except for the MM-B1 cells with 40 50% of dead cells, MM cells did not exert more than 10 15% cell death at pharmacologically relevant doses of TRAIL.
Cells were incubated with Dihydrorhodamine 123 (DHR 123, Sigma Chemical Co., St . Louis MO, USA) for 2 h, washed twice with PBS buffer and subsequently resuspended in propidium iodide solution 0.46 mM for the identification of dead or severely compromised cells.
At high concentration of SNP, such as 0.5 mM for CCL cells and 1 mM for CaCL cells, the proportion of dead cells increases dramatically that we assume cell death might have changed its shape from apoptosis to necrosis.
Interestingly, the morphological view of dead animals by 0.1 mM HWY-4213 was very similar to that by mibefradil but clearly different from that by verpamil (Fig. S3).
The intrathecal catheter consisted of polyethylene tubing that was 12 cm long (PE 10, Intramedic; Clay Adams, Parsippany, NJ) with an outside diameter of 0.4 mm and a dead space of 10 μL that had been sterilized by immersion in 70% (v/v) ethanol and been fully flushed with sterile water before insertion.
The best chamber geometry adopted after optimization was 50 mm flame holder diameter, 60 cm chamber height, having 4 holes of 6, 8 and10 mm with dead zone between the combustion zone and dilution zone.
For Nv odd splice block morpholino, which yielded a high percentage of dead embryos with no cuticle, injections were also carried out at 1 mM, 0.5 mM, and 0.05 mM dilutions.
The percentages of dead cells in a constant area (1.44 mm) were calculated as the number of dead cells/total number of cells × 100; each experiment was repeated 3 5 times.
The total active area of the array is therefore 32×16 mm2 while the total area of the detector (including 1 mm border dead area) is 34×18mm2.
The percentage of dead cells increased in cultures treated with 5 and 10 mM metformin.
