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Inhibition of reactive oxygen species (ROS) production

VJ V. Jaenen
SF S. Fraguas
KB K. Bijnens
MH M. Heleven
TA T. Artois
RR R. Romero
KS K. Smeets
FC F. Cebrià
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The nonspecific flavoprotein inhibitor, Diphenyleneiodonium chloride (DPI, Sigma Aldrich, D2926), was used in order to block ROS production by interfering with several electron transporters. Animals were exposed to 3 μM DPI for 5 h prior to in vivo ROS staining or inflicting a second cut in MEK-inhibited tails, and 1 h prior to amputation when followed by pERK immunohistochemistry. In both cases, animals were continuously exposed to DPI during the regeneration period. Because of its hydrophobic character, DPI was prepared in 0.01% (v/v) dimethylsulfoxide (DMSO, Sigma Aldrich, 471267). In all experiments using DPI, a DMSO-exposed control group was added to take into account the possible effects of DMSO since relatively high concentrations can have neurotoxic effects and influence cell proliferation in S. mediterranea70.

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