Skip to main content
. 2005 Feb;166(2):521–531. doi: 10.1016/S0002-9440(10)62274-2

Figure 6.

Figure 6

Analyses of RT-PCR transcripts and Western blot. Human tau mRNA transcripts (exon 9–11) were detected as 390 bp only in Tg mice brains, but not in non-Tg mice brains, and the intensity of PCR products was at the same level as 6- and 10-month-old Tg mice brains (a, upper panel). Mouse tau mRNA transcripts were detected in both Tg and non-Tg mice brains at the same level at 6 and 10 months of age. The signal intensities of the bands of the human tau mRNA transcripts expressed more than those of mouse tau (a, lower panel). The expression level of human tau R406W in a TS-soluble fraction was relatively higher in cerebral cortical areas, the hippocampus, caudate putamen, thalamus and cerebellum, in addition to the brainstem-spinal cord (b, upper panel). The expression of TS-soluble tau is not observed in non-Tg control mice (b, lower panel). In the TS-soluble fraction of the brain, the expression rate of total tau protein detected by antibody tau-C is 1.8 times higher in the Tg mouse at 6 months of age compared with an age-matched non-Tg mouse (c, upper panel). A Western blot using AT8 showed that tau in a TS-soluble fraction of Tg mice was phosphorylated (c, lower panel). In two 10-month-old TgTau(R406W) brains (Tg1, 21807 and Tg2, 21783), accumulated tauR406W was recognized in the sarcosyl-insoluble fractions and corresponded to the highest 67-kd band of recombinant tau 6 isoforms. Smear tau was detected in the brain of an AD patient; however, no accumulation of sarcosyl-insoluble tau is observed in non-Tg control mice (d). Accumulated tau was detected by PHF-1, Alz-50, and AT8, which corresponded to the highest 67 kd band of recombinant tau 6 isoforms (e, upper panel). Several phosphorylated tau antibodies showed a 67-kd band corresponding to the longest tau isoform, and accumulated tau was phosphorylated at several sites of human tau (PS199, PT205, PT231/PS235, PS396, PS413) (e, middle panel). GSK-3β, especially activated GSK-3β (PY216), showed high accumulation to 48kD in Tg mice brains. On the contrary, PS9, an inactivated GSK-3β antibody, did not show any bands (e, lower panel).