Fig 1.
Microarray analysis of cerebrum specific genes regulated by oxidative stress and aging.
A, Each circle is indicating the numbers of either up or down regulated genes by oxidative stress, aging in the cerebrum and in the cerebellum. Five genes were up-regulated and four genes were down regulated by both oxidative stress and aging in the cerebrum without significant changes in the cerebellum. B, The heat maps of five genes up-regulated and C, four genes down-regulated by oxidative stress and aging. Red indicates up-regulated genes and blue indicates down-regulated genes, while white indicates no significant change in expression. The color gradients indicate the intensity of expression. The data shows the results of four mice per each group. Abbreviation used; OS; oxidative stress.
Fig 2.
Induction of Pla2g3 expression in cerebrum by oxidative stress.
A, B, Quantitative RT-PCR results of Pla2g3 in cerebrum and cerebellum are shown. Fold changes to the aged wild-type mice on normal diet are indicated. n = 4 in each group. C, Western blots for Pla2g3 and alpha-tubulin are shown. Abbreviation used; WT normal; wild type 29 months old mice fed on normal diet, WT deficient; 29 months old wild type mice fed on vitamin E deficient diet, ttpKO deficient; 29 months old Ttpa-/- mice fed on vitamin E deficient diet. *p<0.05.
Fig 3.
Astrocytic expression of Pla2g3 by chronic oxidative stress.
Double immunostaining of Pla2g3 (green) and GFAP (red) in 29 months old wild type mouse (A, B), 29 months old Ttpa-/- mouse (C, D), Ischemic site (E, F) and traumatic injury site (G, H) of cerebral cortex of wild type mice. Arrow heads indicate the strong Pla2g3 expression in astrocytes of Ttpa-/- cortex. Scale bar: 50 μm.
Fig 4.
Overexpression of Pla2g3 reduces IDE expression.
A, B, Quantitative RT-PCR result of IDE in TR-AST cells and rat primary astrocytes. TR-AST cells and primary astrocytes were treated with hydrogen peroxide in indicated conditions. Fold changes to the non-treated cells as controls are indicated. C, Quantitative PCR results of IDE in Pla2g3 transfected HEK293 cells. Human Pla2g3 was transiently expressed and cells were harvested after 48 hours of transfection. Fold changes to the mock control are indicated. *p<0.05, ***p<0.001.
Fig 5.
Increased expression of Pla2g3 in frontal cortex of AD patients.
A, Pla2g3 expression in normal control brain and AD brain. The images are representative of 6 cases in each group. Frontal cortex was stained with anti-Pla2g3 antibody. B, Analysis of Pla2g3-positive cell numbers in frontal cortex. *p<0.05 C-H, Double immunostaining of Pla2g3 and GFAP in normal control brain (C-E) and AD brain (F-H). Arrowheads indicate the intensive staining of Pla2g3 in astrocytes of AD brain. Scale bars: A, 300 μm; C, 50 μm. Abbreviation used; NC; normal control, AD; Alzheimer’s disease patient, I; Molecular layer, II; External granular layer.