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. 2021 Jan 25;13:610037. doi: 10.3389/fnmol.2020.610037

Figure 3.

Figure 3

Specific inhibition of anterograde FAT can be blocked by incubation with a PAD antibody. (A) Pseudophosphorylated tau monomers were incubated with TNT1 antibody which specifically binds the PAD (Kanaan et al., 2011) for 1 h before perfusion into the isolated squid axoplasm. (B) Even after incubation of hTau40-S199E with TNT1 FAT was still inhibited in both directions. (C) Incubation of TNT1 with hTau40-T205E prevented the specific inhibition of anterograde FAT. (D) Co-perfusion of TNT1 with hTau40-S202E/T205E did not prevent inhibition of FAT. (E) Quantitative analysis of anterograde FAT revealed no significant difference between hTau40-S199E perfused with (n = 4) or without (n = 5) incubation with TNT1 (p = 0.223). By contrast, hTau40-T205E + TNT1 (n = 3) was comparable to hTau40-WT (n = 5; p = 0.430) and was significantly different to hTau40-T205E without TNT1 (n = 7; p < 0.001). There was no significant difference between hTau40-S202E/T205E alone (n = 5) or hTau40-S202E/T205E + TNT1 (n = 4; p = 0.466). (F) hTau40-S199E + TNT1 (n = 3) still significantly inhibited retrograde FAT compared to hTau40-WT (n = 5; p = 0.008). hTau40-T205E (n = 7) did not inhibit retrograde FAT and there was no change with TNT1 (n = 3; p = 0.389). hTau40-S202E/T205E + TNT1 (n = 3) was not significantly different from hTau40-S202E/T205E alone (n = 4; p = 0.577). All scatter plots are presented as mean ± 95% confidence interval.