Effects of pertussis toxin on H2O2 generation and receptor autophosphorylation neurons stimulated with insulin. (A) PTX dose–response curve for receptor autophosphorylation in CGN exposed to 100 nM insulin for 10 min (black triangles, mean±SEM of 3 to 8 cultures, *P<0.05 vs. 100 nM insulin). (B) Left Y axis: time courses of H2O2 efflux from CGN exposed to control buffer (white squares, mean of 3 culture dishes), 2 mg/L PTX (light grey squares, mean of 3 culture dishes), 100 nM insulin (red squares, mean of 3 culture dishes), or 100 nM insulin plus 2 mg/L PTX (grey squares, mean of 3 culture dishes). Right Y axis: first time derivatives (rates) of H2O2 efflux from CGN exposed to 100 nM insulin (red line) or 100 nM insulin plus 2 mg/L PTX (black line). (C) Left Y axis: receptor autophosphorylation in CGN exposed to control buffer, 2 mg/L PTX, 100 nM insulin, or 100 nM insulin plus 2 mg/L PTX. Black columns represent the mean±SEM of values obtained from 3 to 8 cultures. *P<0.05 vs. control. #P<0.05 vs. insulin. Right Y axis: Areas under curves (AUC) of H2O2 efflux for 30 s from CGN exposed to control buffer, 2 mg/L PTX, 100 nM insulin, or 100 nM insulin plus 2 mg/L PTX. Red columns represent the mean±SEM of values obtained from 3 culture dishes. *P<0.05 vs. control. #P<0.05 vs. insulin.