Figures
![Figure 1.](/tables/wjon1088w-g001.jpg)
Figure 1. Exposure of B16F10 cells to radiation increased phosphorylated ERK1/2, AKT and JNK levels. There were no significant differences in any of the measured phosphorylated proteins between control and sham-treated groups. Phosphorylated ERK1/2 levels - control or sham versus 1.0 (P < 0.001); control or sham versus 2.0 and 3.0 Gy (P < 0.0001). Phosphorylated AKT levels - control versus 1.0 Gy (P < 0.01); sham versus 1.0 Gy (P < 0.001); control or sham versus 2.0 and 3.0 Gy (P < 0.0001). Phosphorylated JNK levels - control or sham versus 1.0, 2.0 and 3.0 Gy (P < 0.0001).
![Figure 2.](/tables/wjon1088w-g002.jpg)
Figure 2. Cell death in control B16F10 cells, sham-irradiated cells, 2.0 Gy irradiated cells and 3.0 Gy irradiated cells. Compared to control and sham-irradiated cells, 2.0 Gy and 3.0 Gy of irradiation significantly increased cell death (P < 0.0001). Further, compared to 2.0 Gy irradiation, irradiation with 3.0 Gy significantly increased cell death (P < 0.001). Pink color indicates viable cell fraction and green color indicates dead cell fraction.
![Figure 3.](/tables/wjon1088w-g003.jpg)
Figure 3. Effect of inhibition of ERK1/2, AKT and JNK pathways prior to 2.0 Gy radiation exposures of B16F10 cells. B16F10 cells were treated with 20 µM concentrations of U0126, LY294002 and SP600125, 24 h before exposing cells to 2.0 Gy of radiation. Pink color indicates viable cell fraction and green color indicates dead cell fraction. Cell death in U0126 + 2.0 Gy group versus 2.0 Gy alone: P < 0.0001; cell death in LY294002 + 2.0 Gy group versus 2.0 Gy alone: P < 0.0001; cell death in SP600125 + 2.0 Gy group versus 2.0 Gy alone: P < 0.01; cell death in U0126 + LY29400 + 2.0 Gy versus 2.0 Gy alone: P < 0.001.
![Figure 4.](/tables/wjon1088w-g004.jpg)
Figure 4. Increase in amount of cell death (%) with inhibitors of ERK1/2, AKT and JNK pathways. Inhibition of either ERK1/2 or AKT pathways enhanced radiation-induced cell death almost equally (about 43% at 2.0 Gy and about 55% at 3.0 Gy), while inhibition of JNK enhanced relatively low amount of cell death (about 12%) at 2.0 Gy as well as at 3.0 Gy. Combined inhibition of U0126 + LY294002 prior to radiation treatment enhanced radiation induced cell death at 2.0 Gy by 45% and at 3.0 Gy by 51%. Overall no significant difference in percent increase in cell death was observed between U0126, LY294002 or their combination both with 2.0 Gy and 3.0 Gy treatments.
![Figure 5.](/tables/wjon1088w-g005.jpg)
Figure 5. Effect of inhibition of ERK1/2, AKT and JNK pathways prior to 3.0 Gy radiation exposures of B16F10 cells. B16F10 cells were treated with 20 µM concentrations of U0126, LY294002 and SP600125, 24 h before exposing cells to 3.0 Gy of radiation. Pink color indicates viable cell fraction and green color indicates dead cell fraction. Cell death in U0126 + 3.0 Gy group versus 3.0 Gy alone: P < 0.0001; cell death in LY294002 + 3.0 Gy group versus 3.0 Gy alone: P < 0.0001; cell death in SP600125 + 3.0 Gy group versus 3.0 Gy alone: P < 0.01; cell death in U0126 + LY29400 + 3.0 Gy versus 3.0 Gy alone: P < 0.0001.