Fig. 4.

Effect of EP overexpression on p38 MAP kinase activation and mitochondrial dysfunction in Aβ-insulted brain in vivo and brain slices in vitro. a Brain slices from 3-month-old non-Tg or Tg Sh3gl2 mice were perfused with Aβ (50 nM) or vehicle for 1 h and then subjected to immunoblotting analysis for the phosphorylation of p38 MAP kinase (p-p38), total p38 MAP kinase (t-p38), tubulin, and β-actin. Tubulin and β-actin served as a neuronal marker and protein loading controls, respectively. Data are expressed as fold change relative to the non-Tg vehicle control group. b Immunoblotting of cortical homogenates from the indicated Tg mice at 5–6 months of age for the indicated proteins. Data are expressed as fold change relative to the non-Tg mice group. c Brain slices from indicated Tg Sh3gl2 mice were treated with vehicle or Aβ (50 nM) with/without pretreatment of EUK-134 (EUK, 500 nM), SB203580 (SB, 1 µM), or mitochondrial antioxidant MitoTEMPO (TEMPO, 1 µM) for 5 min, and then subjected to immunoblotting for the phosphorylation of p38 MAP kinase (p-P38), total p38 MAP kinase (t-p38), tubulin, and β-actin. Data are expressed as fold change relative to the Tg Sh3gl2 vehicle control group. Date are shown as mean ± s.e.m., n = 3 per group (one-way ANOVA in a–c). d Representative spectra of EPR in non-Tg and Tg Sh3gl2 brain slices with the treatment of vehicle or Aβ (50 nM) in the presence of SB203580 (1 µM). e Quantification of EPR spectra in the indicated groups of mice. f–g Mitochondrial complex IV activity (f) and ATP levels (g) in the indicated groups of brain slices treated with vehicle or Aβ in the presence/absence of SB203580. Data are expressed as fold increase relative to non-Tg vehicle control group. Date are shown as mean ± s.e.m., n = 3 per group (one-way ANOVA in e–g)