Figure 9.

CaD modulates the ROS-NADPH oxidase pathway during monocyte-to-macrophage differentiation and inflammation. THP-1 monocytes were pretreated with CaD (10 µmol/L), or various concentrations (0–10 µmol/L) of CaD/N-acetylcysteine (NAC) for 1 h, followed by PMA (30 nmol/L) treatment for various time points (Ai,Bi), for 4 h (Aii), or 24 h (Bii,C). ROS production (A) was measured using the cell-permeable indicator H₂DCF-DA as described in the Materials and Methods section. NOX2 transcript levels (B) were measured by quantitative RT-PCR, and total p47phox protein expression (T) and membrane translocation (m) (C) were measured by Western blotting. Δ, fold-change normalized to PMA only (n = 3, mean ± SEM. * p < 0.05 vs. no treatment, ^# p < 0.05 vs. PMA only, Student’s t-test (Ai,Bi), or one-way ANOVA (Aii,Bii). The Western blots represent one from at least three independent experiments.