Immunohistochemistry and Western blotting analyses for 4-HNE in iPLA2β-KO mice.
(A‒D) Immunohistochemistry for 4-HNE in the ST of WT mice (A) and iPLA2β-KO mice at 100 weeks (B‒D). (A) Almost no staining is observed in the ST of WT mice at 100 weeks. (B) In the ST of iPLA2β-KO mice at 100 weeks, the increase of 4-HNE is observed mainly in the white matter (small arrows), and the neuropil is also slightly immunostained with 4-HNE, compared with age-matched WT mice (A). (C, D) In high-power fields, some granules or dots strongly positive for 4-HNE are frequently observed (arrowheads in C) in the neuropil of the ST of iPLA2β-KO mice at 100 weeks. A few spheroids are also faintly immunopositive for 4-HNE (arrow in D). Scale bar in (A) represents 100 μm in (A), (B), and 50 μm in (C), (D), respectively. (E) On Western blotting, expression levels of 4-HNE-protein compound are increased in both ST and SN in iPLA2β-KO mice at 100 weeks in comparison with those of age-matched WT mice, with statistical significance (*p < 0.01, Wilcoxon's rank sum test). Data are presented as the ratio of 4-HNE-protein compound to GAPDH (WT mice = 1.0). Each bar represents the mean ± SD.
