NOX2-Dependent Reactive Oxygen Species Regulate Formyl-Peptide Receptor 1-Mediated TrkA Transactivation in SH-SY5Y Cells

Figure 1

FPR1 stimulation induces NOX2 activation. SH-SY5Y cells were serum-starved for 24 hours and (a) stimulated for 2, 5, or 10 minutes with 0.1 μM N-fMLP or (b) preincubated with 100 ng/mL PTX, or with 5 μM cyclosporin H, or (c) with 100 μM apocynin before the stimulation for 5 minutes with 0.1 μM N-fMLP. Sixty micrograms of whole lysates (a, b, and c) was incubated with a phospho-p47^phox(S359)-specific antibody (α-p-p47^phox(S359)), and an anti-tubulin (α-tubulin) antibody was used as a control for protein loading. Band densitometry was evaluated through a scanner equipped with a densitometric workstation. Serum-starved SH-SY5Y cells were (d) stimulated with N-fMLP for increasing time or (e) preincubated with PTX, or cyclosporin H, or apocynin before the stimulation with 0.1 μM N-fMLP for 5 minutes. Detection of ROS was determined by measuring the level of DCF. compared to unstimulated cells. ^§ compared to N-fMLP stimulated cells.

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