Fig. 5

PBT434 improves iron level following MPTP lesion. 12–14 week old male C57BL/6 mice were lesioned using MPTP which resulted in a lesion size 65–70% cell loss by day 21. MPTP lesioned mice were treated with vehicle (VEH) or PBT434 (30 mg/kg/day) from day 1 to day 21. a Brain samples collected at day 21 were sectioned and scanned using laser ablation-inductively coupled plasma-mass spectrometry. Representative images show Fe distribution in normal, unlesioned, wildtype (C57BL6) mouse brain, MPTP lesioned brain and MPTP + PBT434 treated brain. The heat map quantifies the level of iron in the SN, which is indicated by the arrow. b The concentrations of Fe (mg/kg) in the substantia nigra (SN, includes both compacta and reticulata) of the groups of mice were plotted onto a bar graph. MPTP injury causes a significant elevation in Fe in the SN at day 21 (**p < 0.01, one-way ANOVA, Tukey post hoc) compared with unlesioned mice (UL) which was attenuated by PBT434 (* p < 0.05, one-way ANOVA, Tukey post hoc). c PBT434 significantly prevented the MPTP induced elevation of 8-isoprostane within SN as measured by ELISA (*P < 0.05, One-way ANOVA, Tukey Post Hoc). d Western blot was used to measure levels of levels of DJ-1 in the SN. Levels of DJ-1 were significantly elevated with MPTP treatment in the absence of drug (VEH) and significantly further elevated with PBT434 (TP = Total Protein; OD = optical density; ***P < 0.001, One-way ANOVA, Tukey post hoc). The protein ran at the predicted molecular weight (24 kDa) as can be seen by comparing the position of the proteins with the molecular weight ladder on the right of the image. e α-synuclein levels in mice administered MPTP or MPTP + PBT434 (30 mg/kg/day, were compared with unlesioned controls; UL). SN tissue samples were homogenized to form a lysate, which was assayed by Western blot and quantitated by optical density (OD) normalized to total protein (TP, Ponceau). The protein ran at the predicted molecular weight 14 kDa compared to with the molecular weight ladder on the right of the image. In MPTP lesioned mice α-synuclein was significantly elevated by day 21 (**P < 001, one-way ANOVA, Tukey post hoc). α-synuclein protein levels were significantly lower with PBT434 treatment (*P < 0.05, one-way ANOVA, Tukey post hoc) compared with Vehicle treated animals. f Western blot of MPTP lesioned mice showed a significant reduction in levels of ferroportin protein which were decreased 21 days after the lesion (*P < 0.05). The protein ran at slightly less the predicted molecular weight 63 kDa compared to with the molecular weight ladder on the right of the image. Ferroportin protein levels were significantly higher with PBT434 treatment (*P < 0.05) compared with the vehicle treated animals but not different to unlesioned mice (one-way ANOVA, Tukey post hoc)