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. Author manuscript; available in PMC: 2021 May 20.
Published in final edited form as: Cell Rep. 2021 Apr 27;35(4):109037. doi: 10.1016/j.celrep.2021.109037

Figure 1. Tau seeds are aberrantly modified in primary neurons.

Figure 1.

(A) Transmission electron microscopy (TEM) images of wild-type (WT) and P301L tau fibrils. Tau seeds were generated by sonication of preformed tau fibrils. Scale bar, 200 nm.

(B) Coomassie blue staining of WT and P301L tau monomers (M) and seeds (S) prior to addition to neurons. Band intensity was quantified and normalized to WT monomer.

(C) Primary cortical neurons were treated with WT or P301L tau seeds for 0–72 h, followed by immunoblotting analysis with site-specific tau acetylation (ac-K280 and ac-K369) or phosphorylation (p-S262 and p-S356) and total tau antibodies.

(D) Quantification by densitometry shows that P301L tau seed modifications are significantly increased by 24 h compared with WT tau seed (ac-K280 and p-S262). The extent of tau seed modification was normalized to total tau seed level. Error bars indicate SEM; n = 3 biologically independent experiments. p value was determined by unpaired t test. ***p < 0.001.

(E) WT or P301L (PL) tau monomers and seeds were added to 293A cells with or without lipophilic carrier reagent (Lipofectamine 2000) or primary neurons for 2 days, and lysates were analyzed by immunoblotting to detect aberrant tau modifications as described above.