Figure 3

In vitro sensitivity of JVM-2 following treatment with doxorubicin or vincristine in the presence and absence vehicle, AA, EPA or DHA. Cell viability was determined by MTT assay. Percent (%) cell death was determined by Annexin-V/Propidium Iodide duel stain with flow cytometry. Figure 3A illustrates the % cell viability ± SEM of JVM-2 to doxorubicin (0–7.5 μM) in the presence or absence of vehicle, AA 35 μM, EPA 50 μM or DHA 50 μM. Cells pre-treated with AA, EPA or DHA induced significantly greater decreases in cell viability as compared to vehicle when treated with doxorubicin. Figure 3B illustrates the % cell viability ± SEM of JVM-2 to vincristine (0-250 nM) in the presence or absence of vehicle, AA 35 μM, EPA 50 μM or DHA 50 μM. Compared to vehicle, only cells pre-treated with DHA had significantly greater reductions in cell viability when treatment with vincristine. Figure 3C illustrates the % dead cells ± SEM of JVM-2 following pre-treatment with vehicle, AA 35 μM, EPA 50 μM or DHA 50 μM alone and following treatment with 1.5 μM doxorubicin or 100 nM vincristine. Pre-treatment with DHA alone induced significantly greater cell death as compared to vehicle. Compared to vehicle, Cells pre-treated with DHA had significantly greater cell death when treated with doxorubicin or vincristine. Figure 3D provides 2D graphical representations of Annexin-V/PI Plots. ‘Early’ Apoptosis was defined as cells positive for Annexin-V-FITC only. ‘Late’ Apoptosis was defined as cells positive for Annexin-V-FITC and PI. ‘Necrotic’ was defined as cells positive for PI only. Statistical significance was determined by Multiple Comparison Test with Dunnet’s correction (Figure 3A and B) or Tukey’s correction (Figure 3C). α = 0.05, * <0.05, ** < 0.01, *** < 0.001.