Fig. 3

JB activates c-Jun through ROS induction.a. Western blot (cropped) demonstrating 4 h JB exposure results in a strong upregulation of total c-Jun and activation of c-Jun (S63 phosphorylation) in AML cell lines. Lamin is shown as the loading control and the figure is representative of three independent experiments. b. JB induced intracellular ROS in AML cell lines. The bar charts indicate the fold change in median fluorescence intensity compared to untreated controls upon addition of the oxidative stress indicator CM-H2DCFDA, with the elimination of ROS seen when the anti-oxidant NAC is included. Representative flow cytometry plots of ROS measurements are shown above the corresponding bar charts. c. Western blot results (cropped) showing elimination of JB-dependent c-Jun activation by either ROS scavenger or JNKI, representative of three independent experiments. d. cell counts after 24 h incubation showing a combination of ROS scavenger or JNKI with JB treatment reversed JB-induced cell death, displayed as % viability of untreated control. e. DNA damage, assessed by the response marker γH2AX, is increased in JB-treated cells. Bars represent the mean of the Median Fluorescence Intensity (MFI) in respect to the negative untreated control. Etoposide was used as a positive control. Columns, mean of at least three independent experiments; bars, SD. * P < 0.05 and ** P < 0.01. Full length blots for the westerns in this figure are shown in additional file 3