Fig 8.

Part A Effect of KLS-13019 on paclitaxel-induced decreases in fluorescence from the viability dye Alamar Blue during a 5 hour test period on day 9 in culture. A representative experiment is shown in which the protective signal (the RFU [relative fluorescence units] difference between cultures treated with 100 nM KLS-13019 and 3 μM paclitaxel and those treated only with paclitaxel) was 4065 RFU. In contrast, the siRNA signal produced between the difference in fluorescence units from cultures treated with KLS-13019 plus paclitaxel from those treated with mNCX-1 siRNA and KLS-13019 and 3 μM paclitaxel was 3348 RFU. From these data, mNCX-1 siRNA produced a loss of 82 ± 3% of the protective signal. All experiments were conducted with this design, with only the protective agent (10 μM CBD or 100 nM KLS-13019) being different among the studies. In part B, a summary is presented for the comparison of the siRNA-mediated decreases in protective activity produced by either 100 nM KLS-13019 or 10 μM CBD in the dark bars in replicate experiments for each protective agent. In the gray bars, the corresponding changes in neuritic mNCX-1 total spot area/10 fields for each experiment are shown. All bars represent the mean of 6 values ± the standard error. There were no significant differences among either the gray bars or among the dark bars in all 4 experiments indicating a similar % decrease in neuritic target mNCX-1 IR area and the % decrease in protective activity after mNCX-1 siRNA treatment.