Fig. 6
Clomipramine reduces iron neurotoxicity and proliferation of T-lymphocytes and B-lymphocytes. Clomipramine attenuated iron-mediated neurotoxicity in a concentration-dependent manner from 100 nM (p < 0.05) (a). Washing away clomipramine led to cell death by iron, but this effect could be prevented after pre-incubation of clomipramine with iron, suggesting a physical reaction between clomipramine and iron (b). Live-cell imaging studies show that the increasing accumulation of PI-positive neurons exposed to iron over time was prevented by clomipramine (c). Clomipramine furthermore reduced the proliferation of T-lymphocytes (d), reflected by a reduction of cells in S-phase and an increase in the G1-phase of the cell cycle (e, f). Proliferation of activated B-cells was reduced by clomipramine from 2 µM (g), correspondent with reduced TNF-α release (h). Data are shown as quadruplicate replicate wells of an individual experiment that was conducted twice (a, d, e, f), once (b) or three times (g, h); c represents triplicate wells of one experiment. Results are mean ± SEM. One-way ANOVA with Dunnett's multiple comparisons test as post hoc analysis compared to the FeSO4 or activated condition (a, b, d–h) and two-way ANOVA with Dunnett's multiple comparisons test (c): *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001
