Fig 3.

METH-induced superoxide production is mediated by an active NOX complex in hCMEC/D3 cells. (A) hCMEC/D3 cells were treated with 10 μM METH for the indicated time period and the production of superoxide was measured by the oxidation of DHE as determined by flow cytometry. Relative superoxide levels (left) were expressed as percentage of control. (Right) Reflects an example of FACS analysis in hCMEC/D3 cells exposed to METH for 30 min. The arrow indicates a shift of histogram in the METH group indicating increased production of superoxide. (B) Inhibition of NOX protects against METH-induced production of superoxide. hCMEC/D3 cells were pre-treated with 100 μM NSC 23766 (NSC), followed by exposure to 10 μM METH for 30 min. Production of superoxide (left) was quantified by DHE staining and FACS analysis as in (A) (left) and visualized under the confocal microscope (right). Scale bar, 50 μm. The values in (A) and (B) represent the mean ± S.E.M., n= 3; *Statistically significant as compared to the respective control. ^†Values in the METH+NSC group are significantly different as compared to those in the METH group.