Figure 4. Translocation of cytosolic p47^phox to the membrane is prevented by AANO₂.

The effects of AANO₂ on the migration of cytosolic subunits to the membrane were evaluated in activated macrophages. Macrophages (3×10⁶ cells) were treated with AANO₂ (2 and 10 μM) for 15 minutes, washed and PMA-activated as previously. Cells were harvested and membrane (A) as well cytosolic (B) fractions were obtained by differential centrifugation. The presence of p47^phox was determined by western blot. In parallel, the levels of gp91^phox (A) as well as actin (B) were determined as reporters of membrane and cytosolic constitutive proteins, respectively. In (A) the bars represent the relative presence of p47^phox respect to gp91^phox, and results shown correspond to the mean ± SD, n=3. ^& indicates statistical difference compared to the control condition, p < 0.05; * indicates statistical difference compared to the PMA-activated condition, p < 0.05.