Sulforaphane Modulates AQP8-Linked Redox Signalling in Leukemia Cells

Figure 6

Effect of SFN on intracellular ROS level in B1647 cell line after Nox4 inhibition or silencing. (a) B1647 cells were incubated for 24 h with different SFN concentrations, then treated or not with 1 μM plumbagin for 30 min. (b) B1647 cells were transfected with specific siRNA against Nox4 or a random RNA sequence (scrambled) as negative control, C (Scr). 24 h after transfection with siRNA, B1647 cells were incubated for 24 h with different SFN concentrations. Intracellular ROS level was then evaluated as DCF fluorescence as reported in Materials and Methods section. Data are expressed as % of control and represent means ± SD of three independent experiments. Data were analysed by one-way ANOVA followed by Bonferroni’s test. ; ; , significantly different from relative control cells. , significantly different from the corresponding bars in the absence of plumbagin (a) or in Nox4-silenced cells (b). (c) B1647 cells were transfected by electroporation with siRNA against Nox4 or a random RNA sequence (scrambled) as negative control. Effect of RNA interference of Nox4 was confirmed by Western blot analysis with specific antibodies against Nox4.

(a)

(b)

(c)