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Figure 3 | Acta Neuropathologica Communications

Figure 3

From: Relocation of p25α/tubulin polymerization promoting protein from the nucleus to the perinuclear cytoplasm in the oligodendroglia of sporadic and COQ2 mutant multiple system atrophy

Figure 3

TPPP in the nucleus and mitochondria of oligodendroglia. a & b: Immunocryo-ultramicrotomy analysis of an adult mouse brain using TPPP-C-psB. A high magnification view of the square in panel (a) depicts gold particles in both the perinuclear cytoplasm and nucleus (b). c, d: Immunoelectron microscopy of a post-embedded sample using TPPP-N-mab#2A8G3. A high magnification view of a square in the panel (c) demonstrates gold particles again in the cytoplasm and nucleus (d). e, f: Gold particles are not seen on endothelial cells. g, h: Non-specific reactions were not seen on the samples treated with normal goat serum. i: Gold particles were also seen in mitochondria (arrows). a b & j: TPPP-C-psB, c-f & i: TPPP-N-mab#2A8G3. Scale bar = 500 nm (a, d, f, h, i), 100 nm (b), 1 µm (c), 2µm (e, g). j: Western blotting analysis of subcellular fractionation samples from human frontal lobe tissue (Case 26). TPPP-C-psB was used to detect TPPP. Anti-β-tubulin, anti-TOM20, anti-histone H3 antibodies were used as a fractionation control of cytosolic, mitochondrial and nuclear proteins, respectively. The results indicate that TPPP is present not only in the cytosolic, but also in the mitochondrial and nuclear fractions. Considering that comparable amounts of proteins are loaded, the TPPP is contained in the mitochondrial fraction (j). k: The TPPP is not detected in the protein fraction (lane 3) which contains calnexin-positive microsomal proteins, but devoid of mitochondrial (TOM20) and cytosolic (β-tubulin) proteins.

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