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. 2019 Sep 19;13(4):684–699. doi: 10.1016/j.stemcr.2019.08.011

Figure 3.

Figure 3

Reduction of Phosphorylation in R406W Mutant Tau by Multiple Kinases

(A–D) Western blot analysis investigating tau phosphorylation levels at S409 (A) or T181 (C). Both pS409 and pT181 levels were significantly reduced in the R406W mutant samples (S409, B; T181, D) relative to total tau levels (K9JA) (n = 2–3 independent experiments).

(E and F) Western blot analysis with pan-tau antibody Tau5 (E) revealed an increase in the ratio of 48-kDa tau (arrowhead) to total tau (both 48-kDa and 55-kDa bands) in the R406W mutant sample (F) (n = 3–4 independent experiments).

(G and H) Western blot analysis investigating the effects of kinase inhibitors SB216763, KT5720, Y27632, and Roscovitine (inhibitors of GSK3β, PKA, RhoK, and CDK5, respectively) on the phosphorylation pattern of tau in WT (201B7) iPSC-derived neurons (G). Quantification of the ratio of 48-kDa tau to total tau (both 48-kDa and 55-kDa bands) revealed an increase of the ratio with SB216763 treatment, a GSK3β inhibitor, in comparison with that with the mock treatment of DMSO (H) (n = 3 independent experiments).

Error bars indicate mean ± SEM. One-way ANOVA followed by Tukey's test was performed. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. See also Figures S2, S3, and S5.